Compositions and methods for lowering triglycerides in a subject on concomitant statin therapy and having hscrp levels of at least about 2 mg/l

ABSTRACT

In various embodiments, the present disclosure provides compositions and methods for treating and/or preventing cardiovascular-related diseases in subject in need thereof having triglyceride levels of about 200 mg/dL to about 499 mg/dL, and hsCRP levels ≥2.0 mg/L.

PRIORITY CLAIM

This application claims priority to U.S. Provisional Application No.62/637,491 filed on Mar. 2, 2018 and U.S. Provisional Application No.62/730,189 filed on Sep. 12, 2018, the entire contents of each of whichare incorporated herein by reference and relied upon.

BACKGROUND

Cardiovascular disease is one of the leading causes of death in theUnited States and most European countries. It is estimated that over 70million people in the United States alone suffer from a cardiovasculardisease or disorder including but not limited to high blood pressure,coronary heart disease, dyslipidemia, congestive heart failure andstroke.

SUMMARY

In some aspects, the present disclosure provides methods of loweringtriglycerides in a subject on stable statin therapy, the methodcomprising orally administering to the subject daily a pharmaceuticalcomposition comprising about 4 g of ethyl eicosapentaenoate and not morethan about 4% docosahexaenoic acid or its esters, by weight of all fattyacids, for a period of at least about 4 weeks, wherein the subject hasbaseline fasting triglyceride levels of about 200 mg/dl to about 500mg/dl and high-sensitivity C-reactive protein (hsCRP) levels of at leastabout 2 mg/L. In some embodiments, the subject has low-densitylipoprotein cholesterol (LDL-C) levels of about 40 mg/dl to about 100mg/dl prior to said administering.

In some embodiments, the subject has mixed dyslipidemia, chronic kidneydisease, and/or reduced kidney function. In another embodiment, thesubject has coronary heart disease or a coronary heart disease riskequivalent.

In another embodiment, the subject exhibits a reduction in one or moreof: non-high-density lipoprotein cholesterol (non-HDL-C),very-low-density lipoprotein cholesterol (VLDL-C),lipoprotein-associated phospholipase A2 (Lp-PLA₂), apolipoprotein B (ApoB), total cholesterol (TC), high-density lipoprotein cholesterol(HDL-C), very-low-density lipoprotein triglyceride (VLDL-TG), remnantparticle like-cholesterol (RLP-C), or hsCRP levels.

In some embodiments, the LDL-C is not increased in the subject upon 4weeks of said administering. In some embodiments, the LDL-C is reducedin the subject upon 4 weeks of said administering and optionally, thesubject exhibits a reduction in one or more of: non-HDL-C, VLDL-C,Lp-PLA₂, Apo B, TC, HDL-C, VLDL-TG, RLP-C, and hsCRP levels. In yetanother embodiment, the subject exhibits an increase in plasma and/orred blood cell (RBC) levels of EPA.

In yet another embodiment, the subject is determined to have chronickidney disease if the subject has an estimated glomerular filtrationrate (eGFR) of less than about 60 mL/min/1.73 m² for at least about 3months and/or reduced kidney function if the subject has an eGFR of lessthan about 90 mL/min/1.73 m² for at least about 3 months.

In some embodiments, the pharmaceutical composition is present in one ormore dosage units.

These and other embodiments of the present invention will be disclosedin further detail herein below.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the median percent change in lipid/lipoproteins,apolipoproteins, markers of oxidation, and inflammation from baseline toweek 12 for 4 g/day EPA-E vs. placebo in patients having baseline hsCRPlevels ≥2.0 mg/L. Values represent median of difference in percentchange from baseline to week 12 for 4 g/day EPA-E vs placebo. *P<0.0001;†P<0.01; ‡P<0.05; NS, not significant vs placebo.

DETAILED DESCRIPTION

While the present disclosure is capable of being embodied in variousforms, the description below of several embodiments is made with theunderstanding that the present disclosure is to be considered as anexemplification of the invention and is not intended to limit theinvention to the specific embodiments illustrated. Headings are providedfor convenience only and are not to be construed to limit the inventionin any manner. Embodiments illustrated under any heading may be combinedwith embodiments illustrated under any other heading.

The use of numerical values in the various quantitative values specifiedin this application, unless expressly indicated otherwise, are stated asapproximations as though the minimum and maximum values within thestated ranges were both preceded by the word “about.” Also, thedisclosure of ranges is intended as a continuous range including everyvalue between the minimum and maximum values recited as well as anyranges that can be formed by such values. Also disclosed herein are anyand all ratios (and ranges of any such ratios) that can be formed bydividing a disclosed numeric value into any other disclosed numericvalue. Accordingly, the skilled person will appreciate that many suchratios, ranges, and ranges of ratios can be unambiguously derived fromthe numerical values presented herein and in all instances such ratios,ranges, and ranges of ratios represent various embodiments of thepresent disclosure.

Abbreviations used herein include, but are not limited to, Apo B,apolipoprotein B; Apo C-III, apolipoprotein C-III; HDL-C, high-densitylipoprotein cholesterol; hsCRP, high-sensitivity C-reactive protein;ICAM-1, intercellular adhesion molecule-I; IL-6, interleukin-6; ITT,intent-to-treat; LDL-C, low-density lipoprotein cholesterol; Lp-PLA₂,lipoprotein-associated phospholipase A2; non-HDL-C, non-high-densitylipoprotein cholesterol; ox-LDL, oxidized low-density lipoprotein;RLP-C, remnant lipoprotein cholesterol; TC, total cholesterol; TG,triglycerides; VLDL-C, very-low-density lipoprotein cholesterol;VLDL-TG, very-low density lipoprotein triglycerides.

In one embodiment, the disclosure provides a method for treatment and/orprevention of cardiovascular-related diseases. The term“cardiovascular-related disease” herein refers to any disease ordisorder of the heart or blood vessels (i.e. arteries and veins) or anysymptom thereof. Non-limiting examples of cardiovascular-related diseaseand disorders include hypertriglyceridemia, hypercholesterolemia, mixeddyslipidemia, coronary heart disease, vascular disease, stroke,atherosclerosis, arrhythmia, hypertension, myocardial infarction, andother cardiovascular events.

The term “treatment” in relation a given disease or disorder, includes,but is not limited to, inhibiting the disease or disorder, for example,arresting the development of the disease or disorder; relieving thedisease or disorder, for example, causing regression of the disease ordisorder; or relieving a condition caused by or resulting from thedisease or disorder, for example, relieving, preventing or treatingsymptoms of the disease or disorder. The term “prevention” in relationto a given disease or disorder means: preventing the onset of diseasedevelopment if none had occurred, preventing the disease or disorderfrom occurring in a subject that may be predisposed to the disorder ordisease but has not yet been diagnosed as having the disorder ordisease, and/or preventing further disease/disorder development ifalready present.

In one embodiment, the present disclosure provides a method of bloodlipid therapy comprising administering to a subject or subject group inneed thereof a pharmaceutical composition as described herein. Inanother embodiment, the subject or subject group hashypertriglyceridemia, hypercholesterolemia, mixed dyslipidemia and/orvery high triglycerides.

In another embodiment, the subject or subject group being treated inaccordance with the methods of the disclosure has a baselinetriglyceride level (or mean or median baseline triglyceride level in thecase of a subject group), fed or fasting, of about 200 mg/dl to about500 mg/dl. In another embodiment, the subject or subject group has abaseline LDL-C level (or mean or median baseline LDL-C level), despitestable statin therapy, of about 40 mg/dl to about 115 or about 40 toabout 100 mg/dl.

In yet another embodiment, the subject or subject group being treated inaccordance with the methods of the disclosure has elevated baselinehsCRP levels. In some embodiments, the subject or subject group haselevated hsCRP levels of at least about 2 mg/L, at least about 3 mg/L,at least about 4 mg/L, at least about 5 mg/L, at least about 6 mg/L, atleast about 7 mg/L, at least about 8 mg/L, at least about 9 mg/L, atleast about 10 mg/L, at least about 11 mg/L, at least about 12 mg/L, atleast about 13 mg/L, at least about 14 mg/L, at least about 15 mg/L, atleast about 16 mg/L, at least about 17 mg/L, at least about 18 mg/L, atleast about 19 mg/L, at least about 20 mg/L, at least about 21 mg/L, atleast about 22 mg/L, at least about 23 mg/L, at least about 24 mg/L, atleast about 25 mg/L, at least about 26 mg/L, at least about 27 mg/L, atleast about 28 mg/L, at least about 29 mg/L, or at least about 30 mg/L.In another embodiment, the subject or subject group has hsCRP levelsbetween about 2 mg/L to about 10 mg/L, about 3 mg/L to about 15 mg/L,about 5 mg/L to about 30 mg/L, about 4 mg/L to about 20 mg/L, about 10mg/L to about 25 mg/L, about 6 mg/L to about 15 mg/L, about 4 mg/L toabout 25 mg/L, or about 2 mg/L to about 30 mg/L.

In one embodiment, the subject or subject group being treated inaccordance with methods of the disclosure is on concomitant statintherapy, for example atorvastatin, rosuvastatin or simvastatin therapy(with or without ezetimibe). In another embodiment, the subject is onconcomitant stable statin therapy at time of initiation of ultra-pureEPA therapy.

In another embodiment, the subject or subject group being treated inaccordance with methods of the disclosure has a body mass index (BMI ormean BMI) of not more than about 45 kg/m².

In some embodiments, the disclosure provides a method of loweringfasting triglyceride levels in a subject, wherein the subject is onstable statin therapy and has baseline fasting triglycerides of about200 mg/dl to about 500 mg/dl and hsCRP levels of at least about 2 mg/L.In some embodiments, the subject has chronic kidney disease and/orreduced kidney function. In some embodiments, the subject does not havechronic kidney disease and/or reduced kidney function. The term “chronickidney disease” herein means the subject has an estimated glomerularfiltration rate (eGFR) of less than about 60 mL/min/1.73 m² for at leastabout 3 months and the term “reduced kidney function” herein means thesubject has an eGFR of less than about 90 mL/min/1.73 m² for at leastabout 3 months.

In one embodiment, the disclosure provides a method of loweringtriglycerides in a subject on stable statin therapy having baselinefasting triglycerides of about 200 mg/dl to about 500 mg/dl, the methodcomprising administering to the subject a pharmaceutical compositioncomprising about 1 g to about 4 g of EPA (e.g. ultra-pure EPA), whereinupon administering the composition to the subject daily for a period ofabout 12 weeks the subject exhibits at least 10%, at least 15%, at least20%, at least 25%, at least 30%, at least 35%, at least 40%, at least45%, at least 50%, at least 55%, at least 60%, at least 65%, at least70%, or at least 75% lower fasting triglycerides than a control subjectmaintained on stable statin therapy (and optionally placebo matching theultra-pure EPA) without concomitant ultra-pure EPA for a period of about12 weeks, wherein the control subject also has baseline fastingtriglycerides of about 200 mg/dl to about 500 mg/dl. The term “stablestatin therapy” herein means that the subject, subject group, controlsubject or control subject group in question has been taking a stabledaily dose of a statin (e.g. atorvastatin, rosuvastatin or simvastatin)for at least 4 weeks prior to the baseline fasting triglyceridemeasurement (the “qualifying period”). For example, a subject or controlsubject on stable statin therapy would receive a constant daily (i.e.the same dose each day) statin dose for at least 4 weeks immediatelyprior to baseline fasting triglyceride measurement. In one embodiment,the subject's and control subject's LDL-C is maintained between about 40mg/dl and about 115 mg/dl or about 40 mg/dl to about 100 mg/dl duringthe qualifying period. The subject and control subject are thencontinued on their stable statin dose for the 12 week period postbaseline.

In one embodiment, the statin is administered to the subject and thecontrol subject in an amount of about 1 mg to about 500 mg, about 5 mgto about 200 mg, or about 10 mg to about 100 mg, for example about 1 mg,about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg,about 8 mg, about 9 mg, or about 10 mg; about 15 mg, about 20 mg, about25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg,about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about80 mg, about 90 mg, about 100 mg, about 125 mg, about 150 mg, about 175mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, about 425mg, about 450 mg, about 475 mg, or about 500 mg. In another embodiment,the subject (and optionally the control subject) has a baseline LDL-Clevel, despite stable statin therapy, of about 40 mg/dl to about 115mg/dl or about 40 mg/dl to about 100 mg/dl. In another embodiment, thesubject and/or control subject has a body mass index (BMI; or mean BMI)of not more than about 45 kg/m².

In another embodiment, the disclosure provides a method of loweringtriglycerides in a subject group on stable statin therapy having meanbaseline fasting triglycerides of about 200 mg/dl to about 500 mg/dl,the method comprising administering to members of the subject group apharmaceutical composition comprising about 1 g to about 4 g ofultra-pure EPA per day, wherein upon administering the composition tothe members of the subject group daily for a period of about 12 weeksthe subject group exhibits at least 10%, at least 15%, at least 20%, atleast 25%, at least 30%, at least 35%, at least 40%, at least 45%, atleast 50%, at least 55%, at least 60%, at least 65%, at least 70%, atleast 75% lower mean fasting triglycerides than a control subject groupmaintained on stable statin therapy without concomitant ultra-pure EPA(optionally with matching placebo) for a period of about 12 weeks,wherein the control subject group also has mean baseline fastingtriglycerides of about 200 mg/dl to about 500 mg/dl. In a relatedembodiment, the stable statin therapy will be sufficient such that thesubject group has a mean LDL-C level about at least about 40 mg/dl andnot more than about 100 mg/dl or about 40 mg/dl to about 100 mg/dl forthe 4 weeks immediately prior to the baseline fasting triglyceridemeasurement.

In another embodiment, the disclosure provides a method of loweringtriglycerides in subject group on stable statin therapy and having amean baseline fasting triglyceride level of about 200 mg/dl to about 500mg/dl, the method comprising administering to members of the subjectgroup a pharmaceutical composition comprising about 1 g to about 4 g ofultra-pure EPA, wherein upon administering the composition to members ofthe subject group daily for a period of about 12 weeks the subject groupexhibits: (a) at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 55%, at least 60%, at least 65%, at least 70%, at least 75% lowermean fasting triglycerides by comparison with a control subject groupmaintained on stable statin therapy without concomitant ultra-pure EPA(optionally with matching placebo) for a period of about 12 weeks, and(b) no serum LDL-C increase, no statistically significant serum LDL-Cincrease, a serum LDL-C decrease, or the subject is statisticallynon-inferior to the control subjects (statin plus optional placebo) inregard to serum LDL-C elevation) no increase in mean serum LDL-C levelscompared to baseline, wherein the control subject also has mean baselinefasting triglycerides of about 200 mg/dl to about 500 mg/dl.

In another embodiment, the disclosure provides a method of loweringtriglycerides in subject on stable statin therapy and having meanbaseline fasting triglyceride level of about 200 mg/dl to about 500mg/dl, the method comprising administering to the subject apharmaceutical composition comprising about 1 g to about 4 g ofultra-pure EPA, wherein upon administering the composition to thesubject daily for a period of about 12 weeks the subject exhibits (a) atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 55%, atleast 60%, at least 65%, at least 70%, or at least 75% lower fastingtriglycerides by comparison with a control subject maintained on stablestatin therapy without concomitant ultra-pure EPA for a period of about12 weeks and (b) no increase in serum LDL-C levels compared to baseline,wherein the control subject also has baseline fasting triglycerides ofabout 200 mg/dl to about 500 mg/dl.

In another embodiment, the disclosure provides a method of loweringtriglycerides in subject group on stable statin therapy and having meanbaseline fasting triglyceride level of about 200 mg/dl to about 500mg/dl, the method comprising administering to members of the subjectgroup a pharmaceutical composition comprising about 1 g to about 4 g ofultra-pure EPA, wherein upon administering the composition to themembers of the subject group daily for a period of about 12 weeks thesubject group exhibits: (a) at least 10%, at least 15%, at least 20%, atleast 25%, at least 30%, at least 35%, at least 40%, at least 45%, atleast 50%, at least 55%, at least 60%, at least 65%, at least 70%, atleast 75% lower mean fasting triglycerides and (b) at least 5%, at least10%, at least 15%, at least 20%, at least 25%, at least 30%, at least35%, at least 40%, at least 45% or at least 50% lower mean serum LDL-Clevels by comparison with a control subject group maintained on stablestatin therapy without concomitant ultra-pure EPA (optionally withmatching placebo) for a period of about 12 weeks, no serum LDL-Cincrease, no statistically significant serum LDL-C increase, nostatistically significant serum LDL-C increase, a serum LDL-C decrease,or the subject group is statistically non-inferior to the controlsubject group (statin plus optional placebo) in regard to serum LDL-Celevation), wherein the control subject group also has mean baselinefasting triglycerides of about 200 mg/dl to about 500 mg/dl.

In another embodiment, the disclosure provides a method of loweringtriglycerides in subject group on stable statin therapy and having meanbaseline fasting triglyceride level of about 200 mg/dl to about 500mg/dl, the method comprising administering to members of the subjectgroup a pharmaceutical composition comprising about 1 g to about 4 g ofultra-pure EPA, wherein upon administering the composition to themembers of the subject group daily for a period of about 12 weeks thesubject group exhibits (a) at least 10%, at least 15%, at least 20%, atleast 25%, at least 30%, at least 35%, at least 40%, at least 45%, atleast 50%, at least 55%, at least 60%, at least 65%, at least 70%, atleast 75% lower mean fasting triglycerides and (b) at least 5%, at least10%, at least 15%, at least 20%, at least 25%, at least 30%, at least35%, at least 40%, at least 45% or at least 50% lower mean serum LDL-Clevels by comparison with a control subject group maintained on stablestatin therapy without concomitant ultra-pure EPA (optionally withmatching placebo) for a period of about 12 weeks, no serum LDL-Cincrease, no statistically significant serum LDL-C increase, nostatistically significant serum LDL-C increase, a serum LDL-C decrease,or the subject group is statistically non-inferior to the controlsubject group (statin plus optional placebo) in regard to serum LDL-Celevation), wherein the control subject group also has mean baselinefasting triglycerides of about 200 mg/dl to about 500 mg/dl.

In another embodiment, the subject or subject group being treated inaccordance with methods of the disclosure exhibits a fasting baselineabsolute plasma level of free total fatty acid (or mean thereof) notgreater than about 300 nmol/ml, not greater than about 250 nmol/ml, notgreater than about 200 nmol/ml, not greater than about 150 nmol/ml, notgreater than about 100 nmol/ml, or not greater than about 50 nmol/ml.

In another embodiment, the subject or subject group being treated inaccordance with methods of the disclosure exhibits a fasting baselineabsolute plasma level of free EPA (or mean thereof in the case of asubject group) not greater than about 0.70 nmol/ml, not greater thanabout 0.65 nmol/ml, not greater than about 0.60 nmol/ml, not greaterthan about 0.55 nmol/ml, not greater than about 0.50 nmol/ml, notgreater than about 0.45 nmol/ml, or not greater than about 0.40 nmol/ml.In another embodiment, the subject or subject group being treated inaccordance with methods of the disclosure exhibits a baseline fastingplasma level (or mean thereof) of free EPA, expressed as a percentage oftotal free fatty acid, of not more than about 3%, not more than about2.5%, not more than about 2%, not more than about 1.5%, not more thanabout 1%, not more than about 0.75%, not more than about 0.5%, not morethan about 0.25%, not more than about 0.2% or not more than about 0.15%.In one such embodiment, free plasma EPA and/or total fatty acid levelsare determined prior to initiating therapy.

In another embodiment, the subject or subject group being treated inaccordance with methods of the disclosure exhibits a fasting baselineabsolute plasma level of free EPA (or mean thereof) not greater thanabout 1 nmol/ml, not greater than about 0.75 nmol/ml, not greater thanabout 0.50 nmol/ml, not greater than about 0.4 nmol/ml, not greater thanabout 0.35 nmol/ml, or not greater than about 0.30 nmol/ml.

In another embodiment, the subject or subject group being treated inaccordance with methods of the disclosure exhibits a fasting baselineplasma, serum or red blood cell (RBC) membrane EPA level not greaterthan about 150 μg/ml, not greater than about 125 μg/ml, not greater thanabout 100 μg/ml, not greater than about 95 μg/ml, not greater than about75 μg/ml, not greater than about 60 μg/ml, not greater than about 50μg/ml, not greater than about 40 μg/ml, not greater than about 30 μg/ml,or not greater than about 25 μg/ml.

In another embodiment, methods of the present disclosure comprise a stepof measuring the subject's (or subject group's mean) baseline lipidprofile prior to initiating therapy. In another embodiment, methods ofthe disclosure comprise the step of identifying a subject or subjectgroup having one or more of the following: baseline non-HDL-C value (ormean) of about 200 mg/dl to about 400 mg/dl, for example at least about210 mg/dl, at least about 220 mg/dl, at least about 230 mg/dl, at leastabout 240 mg/dl, at least about 250 mg/dl, at least about 260 mg/dl, atleast about 270 mg/dl, at least about 280 mg/dl, at least about 290mg/dl, or at least about 300 mg/dl; baseline total cholesterol value (ormean) of about 250 mg/dl to about 400 mg/dl, for example at least about260 mg/dl, at least about 270 mg/dl, at least about 280 mg/dl or atleast about 290 mg/dl; baseline VLDL-C value (or mean) of about 140mg/dl to about 200 mg/dl, for example at least about 150 mg/dl, at leastabout 160 mg/dl, at least about 170 mg/dl, at least about 180 mg/dl orat least about 190 mg/dl; baseline HDL-C value (or mean) of about 10 toabout 100 mg/dl, for example not more than about 90 mg/dl not, not morethan about 80 mg/dl, not more than about 70 mg/dl, not more than about60 mg/dl, not more than about 60 mg/dl, not more than about 50 mg/dl,not more than about 40 mg/dl, not more than about 35 mg/dl, not morethan about 30 mg/dl, not more than about 25 mg/dl, not more than about20 mg/dl, or not more than about 15 mg/dl; and/or baseline LDL-C value(or mean) of about 30 to about 300 mg/dl, for example not less thanabout 40 mg/dl, not less than about 50 mg/dl, not less than about 60mg/dl, not less than about 70 mg/dl, not less than about 90 mg/dl or notless than about 90 mg/dl.

In a related embodiment, upon treatment in accordance with the presentdisclosure, for example over a period of about 1 to about 200 weeks,about 1 to about 100 weeks, about 1 to about 80 weeks, about 1 to about50 weeks, about 1 to about 40 weeks, about 1 to about 20 weeks, about 1to about 15 weeks, about 1 to about 12 weeks, about 1 to about 10 weeks,about 1 to about 5 weeks, about 1 to about 2 weeks or about 1 week, thesubject or subject group exhibits one or more of the following outcomes:

(a) a reduction in triglyceride levels compared to baseline or placebocontrol (e.g., a subject on stable statin plus placebo matching the EPAtreatment group);

(b) a reduction in Apo B levels compared to baseline or placebo control;

(c) an increase HDL-C levels compared to baseline or placebo control;

(d) substantially no change (e.g., no increase), no statisticallysignificant increase, or a reduction in LDL-C levels compared tobaseline or placebo control;

(e) substantially no change (e.g., no increase), no statisticallysignificant increase, or a reduction in non-HDL-C levels compared tobaseline or placebo control;

(f) a reduction in VLDL levels compared to baseline or placebo control;

(g) an increase in apolipoprotein A-I (Apo A-I) levels compared tobaseline or placebo control;

(h) an increase in apo A-I/apo B ratio compared to baseline or placebocontrol;

(i) a reduction in lipoprotein (a) levels compared to baseline orplacebo control;

(i) a reduction in (1) mean LDL particle number (2) total LDL particlenumber, and/or (3) LDL particle number compared to baseline or placebocontrol;

(k) an increase in (1) mean LDL particle size (2) total LDL particlesize, and/or (3) LDL particle size compared to baseline or placebocontrol;

(l) a reduction in remnant-like particle cholesterol (RLP-C) compared tobaseline or placebo control;

(m) a reduction in oxidized LDL compared to baseline or placebo control;

(n) substantially no change (e.g., no increase), no statisticallysignificant change, or a reduction in fasting plasma glucose (FPG)compared to baseline or placebo control;

(o) a reduction in hemoglobin A_(1c) (HbA_(1c)) compared to baseline orplacebo control;

(p) a reduction in homeostasis model insulin resistance compared tobaseline or placebo control;

(q) a reduction in lipoprotein associated phospholipase A2 (Lp-PLA₂)compared to baseline or placebo control;

(r) a reduction in ICAM-1 compared to baseline or placebo control;

(s) a reduction in IL-6 compared to baseline or placebo control;

(t) a reduction in plasminogen activator inhibitor-1 (PAI-1) compared tobaseline or placebo control;

(u) a reduction in hsCRP compared to baseline or placebo control;

(v) an increase in serum, plasma, and/or RBC EPA levels compared tobaseline or placebo control;

(w) an increase in serum phospholipid and/or RBC membrane EPA levelscompared to baseline or placebo control;

(x) substantially no change (e.g., no increase), no statisticallysignificant change, or a reduction in one or more of serum and/or RBCcontent of docosahexaenoic acid (DHA), docosapentaenoic acid (DPA),arachidonic acid (AA), palmitic acid (PA), stearidonic acid (SA) oroleic acid (OA) compared to baseline or placebo control;

(y) an increase in one or more of serum and/or RBC content of DHA, DPA,AA, PA, SA, and/or OA compared to baseline or placebo control;

(z) a reduction in total cholesterol as compared to baseline or placebocontrol;

(aa) a reduction in or prevention of membrane cholesterol domainformation as compared to baseline or placebo control;

(bb) a reduction in or prevention of oxidative modification of membranepolyunsaturated fatty acids as compared to baseline or placebo control;

(cc) no increase, no substantial increase, or a reduction in oxidizedsdLDL as compared to baseline or placebo control;

(dd) an increase in a ratio of EPA to AA levels in serum and/or RBCs ascompared to baseline or placebo control;

(ee) an increase in a ratio of OA to SA levels in serum and/or RBCs ascompared to baseline or placebo control;

(ff) a reduction in a ratio of OA to SA levels in serum and/or RBCs ascompared to baseline or placebo control;

(gg) an increase in a ratio of total omega-3 fatty acids to totalomega-6 fatty acids as compared to baseline or placebo control;

(hh) a reduction in VLDL-TG levels as compared to baseline or placebocontrol; and/or

(ii) a reduction in apolipoprotein C-III (Apo C-III) levels as comparedto baseline or placebo control;

In one embodiment, upon administering a composition of the disclosure toa subject, the subject exhibits a decrease in triglyceride levels, anincrease in the concentrations of EPA and DPA (n−3) in RBCs, and anincrease of the ratio of EPA: arachidonic acid in RBCs. In a relatedembodiment the subject exhibits substantially no or no increase in RBCDHA.

In one embodiment, methods of the present disclosure comprise measuringbaseline levels of one or more markers set forth in (a)-(ii) above priorto dosing the subject or subject group. In another embodiment, themethods comprise administering a composition as disclosed herein to thesubject after baseline levels of one or more markers set forth in(a)-(ii) are determined, and subsequently taking an additionalmeasurement of said one or more markers.

In another embodiment, upon treatment with a composition of the presentdisclosure, for example over a period of about 1 to about 200 weeks,about 1 to about 100 weeks, about 1 to about 80 weeks, about 1 to about50 weeks, about 1 to about 40 weeks, about 1 to about 20 weeks, about 1to about 15 weeks, about 1 to about 12 weeks, about 1 to about 10 weeks,about 1 to about 5 weeks, about 1 to about 2 weeks or about 1 week, thesubject or subject group exhibits any 2 or more of, any 3 or more of,any 4 or more of, any 5 or more of, any 6 or more of, any 7 or more of,any 8 or more of, any 9 or more of, any 10 or more of, any 11 or moreof, any 12 or more of, any 13 or more of, any 14 or more of, any 15 ormore of, any 16 or more of, any 17 or more of, any 18 or more of, any 19or more of, any 20 or more of, any 21 or more of, any 22 or more of, any23 or more, any 24 or more, or all 25 of outcomes (a)-(ii) describedimmediately above.

In another embodiment, upon treatment with a composition of the presentdisclosure, the subject or subject group exhibits one or more of thefollowing outcomes:

(a) a reduction in triglyceride levels of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55% or at least about 75%(actual % change or median % change) as compared to baseline or placebocontrol (e.g. a subject on statin and placebo matching the EPA treatmentgroup);

(b) a reduction in Apo B levels of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, at least about 55% or at least about 75%(actual % change or median % change) as compared to baseline or placebocontrol;

(c) substantially no change, no statistically significant change, or anincrease in HDL-C levels of at least about 5%, at least about 10%, atleast about 15%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at least about 45%, atleast about 50%, at least about 55% or at least about 75% (actual %change or median % change) as compared to baseline or placebo control;

(d) substantially no change (e.g., no increase), no statisticallysignificant increase, or a reduction in LDL-C level of less than 60%increase, less than 50% increase, less than 40% increase, less than 30%increase, less than 20% increase, less than 10% increase, less than 5%(actual % change or median % change) as compared to baseline or placebocontrol;

(e) substantially no change (e.g., no increase), no statisticallysignificant increase, or a reduction in non-HDL-C levels of a less than30% increase, less than 20% increase, less than 10% increase, less than5% (actual % change or median % change) as compared to baseline orplacebo control;

(f) a reduction in VLDL levels of at least about 5%, at least about 10%,at least about 15%, at least about 20%, at least about 25%, at leastabout 30%, at least about 35%, at least about 40%, at least about 45%,at least about 50%, or at least about 100% (actual % change or median %change) compared to baseline or placebo control;

(g) an increase in apo A-I levels of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, or at least about 100% (actual % change ormedian % change) compared to baseline or placebo control;

(h) an increase in apo A-I/apo B ratio of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, or at least about 100% (actual % changeor median % change) compared to baseline or placebo control;

(i) a reduction in lipoprotein (a) levels of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, or at least about 100% (actual % changeor median % change) compared to baseline or placebo control;

(j) a reduction in (1) mean LDL number, (2) total LDL particle, and/or(3) LDL particle number of at least about 5%, at least about 10%, atleast about 15%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at least about 45%, atleast about 50%, or at least about 100% (actual % change or median %change) compared to baseline or placebo control;

(k) an increase in (l) mean LDL particle size, (2) total LDL particlesize, and/or (3) LDL particle size of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, or at least about 100% (actual % change ormedian % change) compared to baseline or placebo control;

(l) a reduction in RLP-C of at least about 5%, at least about 10%, atleast about 15%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at least about 45%, atleast about 50%, or at least about 100% (actual % change or median %change) compared to baseline or placebo control;

(m) a reduction in oxidized LDL of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, or at least about 100% (actual % change ormedian % change) compared to baseline or placebo control;

(n) substantially no change (e.g., no increase), no statisticallysignificant change, or a reduction in FPG of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, or at least about 100% (actual % changeor median % change) compared to baseline or placebo control;

(o) substantially no change, no statistically significant change, areduction in HbA_(1c) of at least about 5%, at least about 10%, at leastabout 15%, at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, or at leastabout 50% (actual % change or median % change) compared to baseline orplacebo control;

(p) a reduction in homeostasis model index insulin resistance of atleast about 5%, at least about 10%, at least about 15%, at least about20%, at least about 25%, at least about 30%, at least about 35%, atleast about 40%, at least about 45%, at least about 50%, or at leastabout 100% (actual % change or median % change) compared to baseline orplacebo control;

(q) a reduction in LpPLA₂ of at least about 5%, at least about 10%, atleast about 15%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at least about 45%, atleast about 50%, or at least about 100% (actual % change or median %change) compared to baseline or placebo control;

(r) a reduction in ICAM-1 of at least about 5%, at least about 10%, atleast about 15%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at least about 45%, atleast about 50%, or at least about 100% (actual % change or median %change) compared to baseline or placebo control;

(s) a reduction in IL-6 of at least about 5%, at least about 10%, atleast about 15%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at least about 45%, atleast about 50%, or at least about 100% (actual % change or median %change) compared to baseline or placebo control;

(t) a reduction in PAI-1 of at least about 5%, at least about 10%, atleast about 15%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at least about 45%, atleast about 50%, or at least about 100% (actual % change or median %change) compared to baseline or placebo control;

(u) a reduction in hsCRP of at least about 5%, at least about 10%, atleast about 15%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at least about 45%, atleast about 50%, or at least about 100% (actual % change or median %change) compared to baseline or placebo control;

(v) an increase in serum, plasma and/or RBC EPA levels of at least about5%, at least about 10%, at least about 15%, at least about 20%, at leastabout 25%, at least about 30%, at least about 35%, at least about 40%,at least about 45%, at least about 50%, at least about 100%, at leastabout 200% or at least about 400% (actual % change or median % change)compared to baseline or placebo control;

(w) an increase in serum phospholipid and/or RBC membrane EPA levels ofat least about 5%, at least about 10%, at least about 15%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, at least about 50%, at leastabout 100%, at least about 200%, or at least about 400% (actual % changeor median % change) compared to baseline or placebo control;

(x) substantially no change (e.g., no increase), no statisticallysignificant change, or a reduction in one or more of serum phospholipidand/or RBC DHA, DPA, AA, PA and/or OA of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55% or at least about 75%(actual % change or median % change) compared to baseline or placebocontrol;

(y) an increase in one or more of serum phospholipid and/or RBC DHA,DPA, AA, PA and/or OA of at least about 5%, at least about 10%, at leastabout 15%, at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at leastabout 50%, at least about 55% or at least about 75% (actual % change ormedian % change) compared to baseline or placebo control;

(z) a reduction in total cholesterol of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55% or at least about 75%(actual % change or median % change) compared to baseline or placebocontrol;

(aa) a reduction in or prevention of membrane cholesterol domainformation of at least about 10%, at least about 15%, at least about 20%,at least about 25%, at least about 30%, at least about 35%, at leastabout 40%, at least about 45%, at least about 50%, at least about 55%,at least about 60%, at least about 65%, at least about 70%, at leastabout 75%, at least about 80%, at least about 85%, at least about 90%,at least about 95%, at least about 98%, at least about 99%, or about100% (actual % change or median % change) as compared to baseline orplacebo control;

(bb) a reduction in or prevention of oxidative modification of membranepolyunsaturated fatty acids of at least about 10%, at least about 15%,at least about 20%, at least about 25%, at least about 30%, at leastabout 35%, at least about 40%, at least about 45%, at least about 50%,at least about 55%, at least about 60%, at least about 65%, at leastabout 70%, at least about 75%, at least about 80%, at least about 85%,at least about 90%, at least about 95%, at least about 98%, at leastabout 99%, or about 100% (actual % change or median % change) ascompared to baseline or placebo control;

(cc) no increase, substantially no increase, or a reduction in oxidizedsdLDL of at least about 5%, at least about 10%, at least about 15%, atleast about 20%, at least about 25%, at least about 30%, at least about35%, at least about 40%, at least about 45%, at least about 50%, atleast about 55%, at least about 60%, at least about 65%, at least about70%, at least about 75%, at least about 80%, at least about 85%, atleast about 90%, at least about 95%, or greater than 95% (actual %change or median % change) as compared to baseline or placebo control;

(dd) an increase in a ratio of EPA to AA levels in serum and/or RBCs ofat least about 5%, at least about 10%, at least about 15%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, at least about 50%, at leastabout 55%, at least about 60%, at least about 65%, at least about 70%,at least about 75%, at least about 80%, at least about 85%, at leastabout 90%, at least about 95%, or greater than 95% (actual % change ormedian % change) as compared to baseline or placebo control;

(ee) an increase in a ratio of OA to SA levels in serum and/or RBCs ofat least about 5%, at least about 10%, at least about 15%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, at least about 50%, at leastabout 55%, at least about 60%, at least about 65%, at least about 70%,at least about 75%, at least about 80%, at least about 85%, at leastabout 90%, at least about 95%, or greater than 95% (actual % change ormedian % change) as compared to baseline or placebo control;

(ff) a reduction in a ratio of OA to SA levels in serum and/or RBCs ofat least about 5%, at least about 10%, at least about 15%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, at least about 50%, at leastabout 55%, at least about 60%, at least about 65%, at least about 70%,at least about 75%, at least about 80%, at least about 85%, at leastabout 90%, at least about 95%, or greater than 95% (actual % change ormedian % change) as compared to baseline or placebo control;

(gg) an increase in a ratio of total omega-3 fatty acids to totalomega-6 fatty acids of at least about 5%, at least about 10%, at leastabout 15%, at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at leastabout 50%, at least about 55%, at least about 60%, at least about 65%,at least about 70%, at least about 75%, at least about 80%, at leastabout 85%, at least about 90%, at least about 95%, or greater than 95%(actual % change or median % change) as compared to baseline or placebocontrol;

(hh) a reduction in VLDL-TG levels of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, at least about 55%, at least about 60%, atleast about 65%, at least about 70%, at least about 75%, at least about80%, at least about 85%, at least about 90%, at least about 95%, orgreater than 95% (actual % change or median % change) as compared tobaseline or placebo control; and/or

(ii) a reduction in Apo C-III levels of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55%, at least about 60%,at least about 65%, at least about 70%, at least about 75%, at leastabout 80%, at least about 85%, at least about 90%, at least about 95%,or greater than 95% (actual % change or median % change) as compared tobaseline or placebo control.

In one embodiment, methods of the present disclosure comprise measuringbaseline levels of one or more markers set forth in (a)-(ii) prior todosing the subject or subject group. In another embodiment, the methodscomprise administering a composition as disclosed herein to the subjectafter baseline levels of one or more markers set forth in (a)-(ii) aredetermined, and subsequently taking a second measurement of the one ormore markers as measured at baseline for comparison thereto.

In another embodiment, upon treatment with a composition of the presentdisclosure, for example over a period of about 1 to about 200 weeks,about 1 to about 100 weeks, about 1 to about 80 weeks, about 1 to about50 weeks, about 1 to about 40 weeks, about 1 to about 20 weeks, about 1to about 15 weeks, about 1 to about 12 weeks, about 1 to about 10 weeks,about 1 to about 5 weeks, about 1 to about 2 weeks or about 1 week, thesubject or subject group exhibits any 2 or more of, any 3 or more of,any 4 or more of, any 5 or more of, any 6 or more of, any 7 or more of,any 8 or more of, any 9 or more of, any 10 or more of, any 11 or moreof, any 12 or more of, any 13 or more of, any 14 or more of, any 15 ormore of, any 16 or more of, any 17 or more of, any 18 or more of, any 19or more of, any 20 or more of, any 21 or more of, any 22 or more of, any23 or more of, any 24 or more of, or all 26 or more of outcomes (a)-(ii)described immediately above.

Parameters (a)-(ii) can be measured in accordance with any clinicallyacceptable methodology. For example, triglycerides, total cholesterol,HDL-C and fasting blood sugar can be sample from serum and analyzedusing standard photometry techniques. VLDL-TG, LDL-C and VLDL-C can becalculated or determined using serum lipoprotein fractionation bypreparative ultracentrifugation and subsequent quantitative analysis byrefractometry or by analytic ultracentrifugal methodology. Apo A1, Apo Band hsCRP can be determined from serum using standard nephelometrytechniques. Lipoprotein (a) can be determined from serum using standardturbidimetric immunoassay techniques. LDL particle number and particlesize can be determined using nuclear magnetic resonance (NMR)spectrometry. Remnants lipoproteins and LDL-phospholipase A2 can bedetermined from EDTA plasma or serum and serum, respectively, usingenzymatic immunoseparation techniques. Oxidized LDL, intercellularadhesion molecule-1 and interleukin-2 levels can be determined fromserum using standard enzyme immunoassay techniques. These techniques aredescribed in detail in standard textbooks, for example TietzFundamentals of Clinical Chemistry, 6^(th) Ed. (Burtis, Ashwood andBorter Eds.), WB Saunders Company.

In one embodiment, subjects fast for up to 12 hours prior to bloodsample collection, for example about 10 hours.

In another embodiment, the subject being treated is in the highest riskcategory of Adult Treatment Panel (ATP) III Classification of LDL,Total, and HDL Cholesterol (mg/dL) (e.g. CHD or CHD Risk Equivalents(10-year risk >20%)). In another embodiment, the subject is in the ATPIII Multiple (2+) risk factor category.

In one embodiment, the disclosure provides a method of loweringtriglycerides in a subject in the highest risk category of AdultTreatment Panel (ATP) III Classification of LDL, Total, and HDLCholesterol (mg/dL) (e.g. CHD or CHD Risk Equivalents (10-yearrisk >20%)). In another embodiment, the subject is in the ATP IIIMultiple (2+) risk factor category. In another embodiment, the methodincludes a step of identifying a subject in the ATP III Multiple (2+)risk factor category prior to administering ultra-pure E-EPA to thesubject.

In another embodiment, the present disclosure provides a method oftreating or preventing primary hypercholesterolemia and/or mixeddyslipidemia (Fredrickson Types IIa and IIb) in a patient in needthereof, comprising administering to the patient one or morecompositions as disclosed herein. In a related embodiment, the presentdisclosure provides a method of reducing triglyceride levels in asubject or subjects when treatment with a statin or niacinextended-release monotherapy is considered inadequate (Frederickson typeIV hyperlipidemia).

In another embodiment, the present invention disclosure a method oftreating or preventing risk of recurrent nonfatal myocardial infarctionin a patient with a history of myocardial infarction, comprisingadministering to the patient one or more compositions as disclosedherein.

In another embodiment, the present disclosure provides a method ofslowing progression of or promoting regression of atheroscleroticdisease in a patient in need thereof, comprising administering to asubject in need thereof one or more compositions as disclosed herein.

In another embodiment, the present disclosure provides a method oftreating or preventing very high serum triglyceride levels (e.g. TypesIV and V hyperlipidemia) in a patient in need thereof, comprisingadministering to the patient one or more compositions as disclosedherein.

In one embodiment, a composition of the disclosure is administered to asubject in an amount sufficient to provide a daily dose of EPA of about1 mg to about 10,000 mg, 25 about 5000 mg, about 50 to about 3000 mg,about 75 mg to about 2500 mg, or about 100 mg to about 1000 mg, forexample about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, about 425mg, about 450 mg, about 475 mg, about 500 mg, about 525 mg, about 550mg, about 575 mg, about 600 mg, about 625 mg, about 650 mg, about 675mg, about 700 mg, about 725 mg, about 750 mg, about 775 mg, about 800mg, about 825 mg, about 850 mg, about 875 mg, about 900 mg, about 925mg, about 950 mg, about 975 mg, about 1000 mg, about 1025 mg, about 1050mg, about 1075 mg, about 1100 mg, about 1025 mg, about 1050 mg, about1075 mg, about 1200 mg, about 1225 mg, about 1250 mg, about 1275 mg,about 1300 mg, about 1325 mg, about 1350 mg, about 1375 mg, about 1400mg, about 1425 mg, about 1450 mg, about 1475 mg, about 1500 mg, about1525 mg, about 1550 mg, about 1575 mg, about 1600 mg, about 1625 mg,about 1650 mg, about 1675 mg, about 1700 mg, about 1725 mg, about 1750mg, about 1775 mg, about 1800 mg, about 1825 mg, about 1850 mg, about1875 mg, about 1900 mg, about 1925 mg, about 1950 mg, about 1975 mg,about 2000 mg, about 2025 mg, about 2050 mg, about 2075 mg, about 2100mg, about 2125 mg, about 2150 mg, about 2175 mg, about 2200 mg, about2225 mg, about 2250 mg, about 2275 mg, about 2300 mg, about 2325 mg,about 2350 mg, about 2375 mg, about 2400 mg, about 2425 mg, about 2450mg, about 2475 mg or about 2500 mg.

In another embodiment, any of the methods disclosed herein are used intreatment of a subject or subjects that consume a traditional Westerndiet. In one embodiment, the methods of the disclosure include a step ofidentifying a subject as a Western diet consumer or prudent dietconsumer and then treating the subject if the subject is deemed aWestern diet consumer. The term “Western diet” herein refers generallyto a typical diet consisting of, by percentage of total calories, about45% to about 50% carbohydrate, about 35% to about 40% fat, and about 10%to about 15% protein. A Western diet may alternately or additionally becharacterized by relatively high intakes of red and processed meats,sweets, refined grains, and desserts, for example more than 50%, morethan 60% or more or 70% of total calories come from these sources.

In another embodiment, any of the methods disclosed herein are used intreatment of a subject or subjects that consume less than (actual oraverage) about 150 g, less than about 125 g, less than about 100 g, lessthan about 75 g, less than about 50 g, less than about 45 g, less thanabout 40 g, less than about 35 g, less than about 30 g, less than about25 g, less than about 20 g or less than about 15 g of fish per day.

In another embodiment, any of the methods disclosed herein are used intreatment of a subject or subjects that consume less than (actual oraverage) about 10 g, less than about 9 g, less than about 8 g, less thanabout 7 g, less than about 6 g, less than about 5 g, less than about 4g, less than about 3 g, less than about 2 g per day of omega-3 fattyacids from dietary sources.

In another embodiment, any of the methods disclosed herein are used intreatment of a subject or subjects that consume less than (actual oraverage) about 2.5 g, less than about 2 g, less than about 1.5 g, lessthan about 1 g, less than about 0.5 g, less than about 0.25 g, or lessthan about 0.2 g per day of EPA and DHA (combined) from dietary sources.

In one embodiment, compositions useful in various embodiments of thedisclosure comprise a polyunsaturated fatty acid as an activeingredient. In another embodiment, such compositions comprise EPA as anactive ingredient. The term “EPA” as used herein refers toeicosapentaenoic acid (e.g. eicosa-5,8,11,14,17-pentaenoic acid) and/ora pharmaceutically acceptable ester, derivative, conjugate or saltthereof, or mixtures of any of the foregoing.

In one embodiment, the EPA comprises all-ciseicosa-5,8,11,14,17-pentaenoic acid. In another embodiment, the EPA isin the form of an eicosapentaenoic acid ester. In another embodiment,the EPA comprises a C₁-C₅ alkyl ester of EPA. In another embodiment, theEPA comprises eicosapentaenoic acid ethyl ester, eicosapentaenoic acidmethyl ester, eicosapentaenoic acid propyl ester, or eicosapentaenoicacid butyl ester. In still another embodiment, the EPA comprises all-ciseicosa-5,8,11,14,17-pentaenoic acid ethyl ester.

In still other embodiments, the EPA comprises ethyl-EPA, lithium EPA,mono, di- or triglyceride EPA or any other ester or salt of EPA, or thefree acid form of EPA. The EPA may also be in the form of a2-substituted derivative or other derivative which slows down its rateof oxidation but does not otherwise change its biological action to anysubstantial degree.

The term “pharmaceutically acceptable” in the present context means thatthe substance in question does not produce unacceptable toxicity to thesubject or interaction with other components of the composition.

In one embodiment, EPA present in a composition suitable for useaccording to the disclosure comprises ultra-pure EPA. The term“ultra-pure” as used herein with respect to EPA refers to a compositioncomprising at least 96% by weight EPA (as the term “EPA” is defined andexemplified herein). Ultra-pure EPA can comprise even higher purity EPA,for example at least 97% by weight EPA, at least 98% by weight EPA or atleast 99% by weight EPA, wherein the EPA is any form of EPA as set forthherein. Ultra-pure EPA can further be defined (e.g. impurity profile) byany of the description of EPA provided herein.

In some embodiments, EPA is present in a composition in an amount ofabout 50 mg to about 5000 mg, about 75 mg to about 2500 mg, or about 100mg to about 1000 mg, for example about 75 mg, about 100 mg, about 125mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375mg, about 400 mg, about 425 mg, about 450 mg, about 475 mg, about 500mg, about 525 mg, about 550 mg, about 575 mg, about 600 mg, about 625mg, about 650 mg, about 675 mg, about 700 mg, about 725 mg, about 750mg, about 775 mg, about 800 mg, about 825 mg, about 850 mg, about 875mg, about 900 mg, about 925 mg, about 950 mg, about 975 mg, about 1000mg, about 1025 mg, about 1050 mg, about 1075 mg, about 1100 mg, about1025 mg, about 1050 mg, about 1075 mg, about 1200 mg, about 1225 mg,about 1250 mg, about 1275 mg, about 1300 mg, about 1325 mg, about 1350mg, about 1375 mg, about 1400 mg, about 1425 mg, about 1450 mg, about1475 mg, about 1500 mg, about 1525 mg, about 1550 mg, about 1575 mg,about 1600 mg, about 1625 mg, about 1650 mg, about 1675 mg, about 1700mg, about 1725 mg, about 1750 mg, about 1775 mg, about 1800 mg, about1825 mg, about 1850 mg, about 1875 mg, about 1900 mg, about 1925 mg,about 1950 mg, about 1975 mg, about 2000 mg, about 2025 mg, about 2050mg, about 2075 mg, about 2100 mg, about 2125 mg, about 2150 mg, about2175 mg, about 2200 mg, about 2225 mg, about 2250 mg, about 2275 mg,about 2300 mg, about 2325 mg, about 2350 mg, about 2375 mg, about 2400mg, about 2425 mg, about 2450 mg, about 2475 mg, or about 2500 mg.

In various embodiments, one or more antioxidants can be present in theEPA (e.g. E-EPA or ultra pure E-EPA). Non-limiting examples of suitableantioxidants include tocopherol, lecithin, citric acid and/or ascorbicacid. One or more antioxidants, if desired, are typically present in theEPA in an amount of about 0.01% to about 0.1%, by weight, or about0.025% to about 0.05%, by weight.

In one embodiment, a composition of the disclosure contains not morethan about 10%, not more than about 9%, not more than about 8%, not morethan about 7%, not more than about 6%, not more than about 5%, not morethan about 4%, not more than about 3%, not more than about 2%, not morethan about 1%, or not more than about 0.5%, by weight of total fattyacids, docosahexaenoic acid or derivative thereof such as E-DHA, if any.In another embodiment, a composition of the disclosure containssubstantially no docosahexaenoic acid or derivative thereof such asE-DHA. In still another embodiment, a composition of the disclosurecontains no docosahexaenoic acid or E-DHA.

In another embodiment, EPA represents at least about 60%, at least about70%, at least about 80%, at least about 90%, at least about 95%, atleast about 97%, at least about 98%, at least about 99%, or 100%, byweight, of all fatty acids present in a composition useful in accordancewith the disclosure.

In another embodiment, a composition of the disclosure contains lessthan 30%, less than 20%, less than 10%, less than 9%, less than 8%, lessthan 7%, less than 6%, less than 5%, less than 4%, less than 3%, lessthan 2%, less than 1%, less than 0.5% or less than 0.25%, by weight ofthe total composition or by weight of the total fatty acid content, ofany fatty acid other than EPA, or derivative thereof. Illustrativeexamples of a “fatty acid other than EPA” include linolenic acid (LA) orderivative thereof such as ethyl-linolenic acid, AA or derivativethereof such as ethyl-AA, DHA or derivative thereof such as ethyl-DHA,alpha-linolenic acid (ALA) or derivative thereof such as ethyl-ALA,stearadonic acid (STA) or derivative thereof such as ethyl-SA,eicosatrienoic acid (ETA) or derivative thereof such as ethyl-ETA and/orDPA or derivative thereof such as ethyl-DPA.

In another embodiment, a composition of the disclosure has one or moreof the following features: (a) eicosapentaenoic acid ethyl esterrepresents at least 96%, at least 97%, or at least 98%, by weight, ofall fatty acids present in the composition; (b) the composition containsnot more than 4%, not more than 3%, or not more than 2%, by weight, oftotal fatty acids other than eicosapentaenoic acid ethyl ester; (c) thecomposition contains not more than 0.6%, 0.5%, 0.4% or 0.3% of anyindividual fatty acid other than eicosapentaenoic acid ethyl ester; (d)the composition has a refractive index (20° C.) of about 1 to about 2,about 1.2 to about 1.8 or about 1.4 to about 1.5; (e) the compositionhas a specific gravity (20° C.) of about 0.8 to about 1.0, about 0.85 toabout 0.95 or about 0.9 to about 0.92; (f) the composition contains notmore than 20 ppm, 15 ppm or 10 ppm heavy metals, (g) the compositioncontains not more than 5 ppm, 4 ppm, 3 ppm, or 2 ppm arsenic, and/or (h)the composition has a peroxide value not more than 5, 4, 3, or 2 Meq/kg.

In another embodiment, a composition useful in accordance with thedisclosure comprises, consists essentially of or consists of at least95% by weight ethyl eicosapentaenoate (EPA-E), about 0.2% to about 0.5%by weight ethyl octadecatetraenoate (ODTA-E), about 0.05% to about 0.25%by weight ethyl nonaecapentaenoate (NDPA-E), about 0.2% to about 0.45%by weight ethyl arachidonate (AA-E), about 0.3% to about 0.5% by weightethyl eicosatetraenoate (ETA-E), and about 0.05% to about 0.32% ethylheneicosapentaenoate (HPA-E). In another embodiment, the composition ispresent in a capsule shell. In still another embodiment, the capsuleshell contains no chemically modified gelatin.

In another embodiment, compositions useful in accordance with thedisclosure comprise, consist essentially of, or consist of at least 95%,96% or 97%, by weight, ethyl eicosapentaenoate, about 0.2% to about 0.5%by weight ODTA-E, about 0.05% to about 0.25% by weight ethyl NDPA-E,about 0.2% to about 0.45% by weight AA-E, about 0.3% to about 0.5% byweight ETA-E, and about 0.05% to about 0.32% by weight HPA-E.Optionally, the composition contains not more than about 0.06%, about0.05%, or about 0.04%, by weight, DHA or derivative thereof such asethyl-DHA. In one embodiment the composition contains substantially noor no amount of DHA or derivative thereof such as ethyl-DHA. Thecomposition further optionally comprises one or more antioxidants (e.g.tocopherol) in an amount of not more than about 0.5% or not more than0.05%. In another embodiment, the composition comprises about 0.05% toabout 0.4%, for example about 0.2% by weight tocopherol. In anotherembodiment, about 500 mg to about 1 g of the composition is provided ina capsule shell. In another embodiment, the capsule shell contains nochemically modified gelatin.

In another embodiment, compositions useful in accordance with thedisclosure comprise, consist essentially of, or consist of at least 96%by weight ethyl eicosapentaenoate, about 0.22% to about 0.4% by weightODTA-E, about 0.075% to about 0.20% by weight NDPA-E, about 0.25% toabout 0.40% by weight AA-E, about 0.3% to about 0.4% by weight ETA-E andabout 0.075% to about 0.25% by weight HPA-E. Optionally, the compositioncontains not more than about 0.06%, about 0.05%, or about 0.04%, byweight, DHA or derivative thereof such as ethyl-DHA. In one embodimentthe composition contains substantially no or no amount of DHA orderivative thereof such as ethyl-DHA. The composition further optionallycomprises one or more antioxidants (e.g. tocopherol) in an amount of notmore than about 0.5% or not more than 0.05%. In another embodiment, thecomposition comprises about 0.05% to about 0.4%, for example about 0.2%by weight tocopherol. In another embodiment, the disclosure provides adosage form comprising about 500 mg to about 1 g of the foregoingcomposition in a capsule shell. In one embodiment, the dosage form is agel- or liquid-containing capsule and is packaged in blister packages ofabout 1 to about 20 capsules per sheet.

In another embodiment, compositions useful in accordance with thedisclosure comprise, consist essentially of or consist of at least 96%,97% or 98%, by weight, ethyl eicosapentaenoate, about 0.25% to about0.38% by weight ODTA-E, about 0.10% to about 0.15% by weight NDPA-E,about 0.25% to about 0.35% by weight AA-E, about 0.31% to about 0.38% byweight ETA-E, and about 0.08% to about 0.20% by weight HPA-E.Optionally, the composition contains not more than about 0.06%, about0.05%, or about 0.04%, by weight, DHA or derivative thereof such asethyl-DHA. In one embodiment the composition contains substantially noor no amount of DHA or derivative thereof such as ethyl-DHA. Thecomposition further optionally comprises one or more antioxidants (e.g.tocopherol) in an amount of not more than about 0.5% or not more than0.05%. In another embodiment, the composition comprises about 0.05% toabout 0.4%, for example about 0.2% by weight tocopherol. In anotherembodiment, the disclosure provides a dosage form comprising about 500mg to about 1 g of the foregoing composition in a capsule shell. Inanother embodiment, the capsule shell contains no chemically modifiedgelatin.

In another embodiment, a composition as described herein is administeredto a subject once or twice per day. In another embodiment, 1, 2, 3 or 4capsules, each containing about 1 g of a composition as describedherein, are administered to a subject daily. In another embodiment, 1 or2 capsules, each containing about 1 g of a composition as describedherein, are administered to the subject in the morning, for examplebetween about 5 am and about 11 am, and 1 or 2 capsules, each containingabout 1 g of a composition as described herein, are administered to thesubject in the evening, for example between about 5 pm and about 11 pm.

In one embodiment, a subject being treated in accordance with methods ofthe disclosure is not on fibrate or nitrate therapy.

In another embodiment, compositions useful in accordance with methods ofthe disclosure are orally deliverable. The terms “orally deliverable” or“oral administration” herein include any form of delivery of atherapeutic agent or a composition thereof to a subject wherein theagent or composition is placed in the mouth of the subject, whether ornot the agent or composition is swallowed. Thus “oral administration”includes buccal and sublingual as well as esophageal administration. Inone embodiment, the composition is present in a capsule, for example asoft gelatin capsule.

A composition for use in accordance with the disclosure can beformulated as one or more dosage units. The terms “dose unit” and“dosage unit” herein refer to a portion of a pharmaceutical compositionthat contains an amount of a therapeutic agent suitable for a singleadministration to provide a therapeutic effect. Such dosage units may beadministered one to a plurality (i.e. 1 to about 10, 1 to 8, 1 to 6, 1to 4 or 1 to 2) of times per day, or as many times as needed to elicit atherapeutic response.

In another embodiment, the disclosure provides use of any compositiondescribed herein for treating moderate to severe hypertriglyceridemia ina subject in need thereof, comprising: providing a subject having afasting baseline triglyceride level of about 500 mg/dl to about 1500mg/dl and administering to the subject a pharmaceutical composition asdescribed herein. In one embodiment, the composition comprises about 1 gto about 4 g of eicosapentaenoic acid ethyl ester, wherein thecomposition contains substantially no docosahexaenoic acid.

EXAMPLES Example 1. Safety and Efficacy of Ultra-Pure EPA

A multi-center, placebo-controlled, randomized, double-blind, 12-weekstudy is performed to evaluate the efficacy and safety of >96% E-EPA inpatients with fasting triglyceride levels ≥200 mg/dl and <500 mg/dldespite statin therapy (the mean of two qualifying entry values needs tobe >185 mg/dl and at least one of the values needs to be ≥200 mg/dl).The primary objective of the study is to determine the efficacy of >96%E-EPA 2 g daily and 4 g daily, compared to placebo, in lowering fastingTG levels in patients with high risk for cardiovascular disease and withfasting TG levels 2200 mg/dL and <500 mg/dL, despite treatment to LDL-Cgoal on statin therapy.

The secondary objectives of this study are the following:

-   -   1. To determine the safety and tolerability of >96% E-EPA 2 g        daily and 4 g daily;    -   2. To determine the effect of >96% E-EPA on lipid and        apolipoprotein profiles including total cholesterol (TC),        non-high-density lipoprotein cholesterol (non-HDL-C), low        density lipoprotein cholesterol (LDL-C), high density        lipoprotein cholesterol (HDL-C), and very high density        lipoprotein cholesterol (VHDL-C);    -   3. To determine the effect of >96% E-EPA (on lipoprotein        associated phospholipase A2 (Lp-PLA₂) from baseline to week 12;    -   4. To determine the effect of >96% E-EPA on low-density        lipoprotein (LDL) particle number and size;    -   5. To determine the effect of >96% E-EPA on oxidized LDL;    -   6. To determine the effect of >96% E-EPA on FPG and HbA_(1c);    -   7. To determine the effect of >96% E-EPA on insulin resistance;    -   8. To determine the effect of >96% E-EPA on hsCRP;    -   9. To determine the effects of >96% E-EPA 2 g daily and 4 g        daily on the incorporation of fatty acids into RBC membranes and        into plasma phospholipids;    -   10. To explore the relationship between baseline fasting TG        levels and the reduction in fasting TG levels; and    -   11. To explore the relationship between changes of fatty acid        concentrations in plasma and RBC membranes, and the reduction in        fasting TG levels.

The population for this study is men and women >18 years of age with abody mass index ≤45 kg/m² with fasting TG levels greater than or equalto 200 mg/dl and less than 500 mg/dl and on a stable does of statintherapy (with or without ezetimibe). The statin must be atorvastatin,rosuvastatin or simvastatin. The dose of statin must be stable for ≥4weeks prior to the LDL-C/TG baseline qualifying measurement forrandomization. The statin dose will be optimal such that the patientsare at their LDL-C goal at the LDL-C/TG baseline qualifyingmeasurements. The same statin at the same dose will be continued untilthe study ends.

Patients taking any additional non-statin, lipid-altering medications(niacin >200 mg/day, fibrates, fish oil, other products containingomega-3 fatty acids, or other herbal products or dietary supplementswith potential lipid-altering effects), either alone or in combinationwith statin therapy (with or without ezetimibe), must be able to safelydiscontinue non-statin, lipid-altering therapy at screening.

Patients at high risk for CVD, i.e., patients with clinical coronaryheart disease (CHD) or clinical CHD risk equivalents (10-year risk >20%)as defined in the National Cholesterol Education Program (NCEP) AdultTreatment Panel III (ATP III) Guidelines will be eligible to participatein this study. Those include patients with any of the followingcriteria: (1) Known CVD, either clinical coronary heart disease (CHD),symptomatic carotid artery disease (CAD), peripheral artery disease(PAD) or abdominal aortic aneurism; or (2) Diabetes Mellitus (Type 1 or2).

Approximately 648 patients will be randomized at approximately 80centers in the U.S. The study will be a 18- to 20-week, Phase 3,multi-center study consisting of 2 study periods: (1) A 6- to 8-weekscreening period that includes a diet and lifestyle stabilization, anon-statin lipid-altering treatment washout, and an LDL-C and TGqualifying period and (2) A 12-week, double-blind, randomized,placebo-controlled treatment period.

During the screening period and double-blind treatment period, allvisits are to be within ±3 days of the scheduled time. All patients willcontinue to take the statin product (with or without ezetimibe) at thesame dose they were taking at screening throughout their participationin the study.

The 6- to 8-week screening period includes a diet and lifestylestabilization, a non-statin lipid-altering treatment washout, and anLDL-C and TG qualifying period. The screening visit (Visit 1) will occurfor all patients at either 6 weeks (for patients on stable statintherapy [(with or without ezetimibe] at screening) or 8 weeks (forpatients who will require washout of their current non-statinlipid-altering therapy at screening) before randomization, as follows:

-   -   Patients who do not require a washout: The screening visit will        occur at Visit 1 (Week −6). Eligible patients will enter a        4-week diet and lifestyle stabilization period. At the screening        visit, all patients will receive counseling regarding the        importance of the National Cholesterol Education Program (NCEP)        Therapeutic Lifestyle Changes (TLC) diet and will receive basic        instructions on how to follow this diet.    -   Patients who will require a washout: The screening visit will        occur at Visit 1 (Week −8). Eligible patients will begin a        6-week washout period at the screening visit (i.e. 6 weeks        washout before the first LDL-C/TG qualifying visit). Patients        will receive counseling regarding the NCEP TLC diet and will        receive basic instructions on how to follow this diet. Site        personnel will contact patients who do not qualify for        participation based on screening laboratory test results to        instruct them to resume their prior lipid-altering medications.

At the end of the 4-week diet and lifestyle stabilization period or the6-week diet and stabilization and washout period, eligible patients willenter the 2-week LDL-C and TG qualifying period and will have theirfasting LDL-C and TG levels measured at Visit 2 (Week −2) and Visit 3(Week −1). Eligible patients must have an average fasting LDL-C level240 mg/dL and <100 mg/dL and an average fasting TG level 2200 mg/dL and<500 mg/dL to enter the 12-week double-blind treatment period. The LDL-Cand TG levels for qualification will be based on the average (arithmeticmean) of the Visit 2 (Week −2) and Visit 3 (Week −1) values. If apatient's average LDL-C and/or TG levels from Visit 2 and Visit 3 falloutside the required range for entry into the study, an additionalfasting lipid profile can be collected 1 week later at Visit 3.1. If athird sample is collected at Visit 3.1, entry into the study will bebased on the average (arithmetic mean) of the values from Visit 3 andVisit 3.1.

After confirmation of qualifying fasting LDL-C and TG values, eligiblepatients will enter a 12-week, randomized, double-blind treatmentperiod. At Visit 4 (Week 0), patients will be randomly assigned to 1 ofthe following treatment groups:

-   -   >96% E-EPA 2 g daily,    -   >96% E-EPA 4 g daily, or    -   Placebo.

Approximately 216 patients per treatment group will be randomized inthis study. Stratification will be by type of statin (atorvastatin,rosuvastatin or simvastatin), the presence of diabetes, and gender.

During the double-blind treatment period, patients will return to thesite at Visit 5 (Week 4), Visit 6 (Week 11), and Visit 7 (Week 12) forefficacy and safety evaluations.

Eligible patients will be randomly assigned at Visit 4 (Week 0) toreceive orally >96% E-EPA 2 g daily, >96% E-EPA 4 g daily, or placebo.

>96% E-EPA is provided in 1 g liquid-filled, oblong, gelatin capsules.The matching placebo capsule is filled with light liquid paraffin andcontains 0 g of >96% E-EPA. >96% E-EPA capsules are to be taken withfood (i.e. with or at the end of a meal).

During the double-blind treatment period, patients will take 2 capsules(>96% E-EPA or matching placebo) in the morning and 2 capsules in theevening for a total of 4 capsules per day.

-   -   Patients in the >96% E-EPA 2 g/day treatment group will receive        1>96% E-EPA 1 g capsule and 1 matching placebo capsule in the        morning and in the evening.    -   Patients in the >96% E-EPA 4 g/day treatment group will receive        2>96% E-EPA 1 g capsules in the morning and evening.

Patients in the placebo group will receive 2 matching placebo capsulesin the morning and evening.

The primary efficacy variable for the double-blind treatment period ispercent change in TG from baseline to Week 12 endpoint. The secondaryefficacy variables for the double-blind treatment period include thefollowing:

-   -   Percent changes in total TC, HDL-C, LDL-C, calculated non-HDL-C,        and VLDL-C from baseline to Week 12 endpoint;    -   Percent change in very low-density lipoprotein TG from baseline        to Week 12;    -   Percent changes in apo A-I, apo B, and apo A-I/apo B ratio from        baseline to Week 12;    -   Percent changes in lipoprotein(a) from baseline to Week 12;    -   Percent changes in LDL particle number and size, measured by        nuclear magnetic resonance, from baseline to Week 12;    -   Percent change in remnant-like particle cholesterol from        baseline to Week 12;    -   Percent change in oxidized LDL from baseline to Week 12;    -   Changes in FPG and HbA_(1c) from baseline to Week 12;    -   Change in insulin resistance, as assessed by the homeostasis        model index insulin resistance, from baseline to Week 12;    -   Percent change in Lp-PLA₂ from baseline to Week 12;    -   Change in intracellular adhesion molecule-1 from baseline to        Week 12;    -   Change in interleukin-2 from baseline to Week 12;    -   Change in plasminogen activator inhibitor-1 from baseline to        Week 12. Note: this parameter will only be collected at sites        with proper storage conditions;    -   Change in hsCRP from baseline to Week 12; and    -   Change in plasma concentration and RBC membrane content of fatty        acid from baseline to Week 12 including EPA, DPA, DHA, AA,        dihomo-γ-linolenic acid (DGLA), the ratio of EPA/AA, ratio of        OA/SA, and the ratio of total omega-3 acids over total omega-6        acids.

Safety assessments will include adverse events, clinical laboratorymeasurements (chemistry, hematology, and urinalysis), 12-leadelectrocardiograms (ECGs), vital signs, and physical examinations.

For TG, TC, HDL-C, LDL-C, calculated non-HDL-C, and VLDL-C, baselinewill be defined as the average of Visit 4 (Week 0) and the precedinglipid qualifying visit (either Visit 3 [Week −1] or if it occurs, Visit3.1) measurements. Baseline for all other efficacy parameters will bethe Visit 4 (Week 0) measurement.

For TG, TC, HDL-C, LDL-C, calculated non-HDL-C, and VLDL-C, Week 12endpoint will be defined as the average of Visit 6 (Week 11) and Visit 7(Week 12) measurements.

Week 12 endpoint for all other efficacy parameters will be the Visit 7(Week 12) measurement.

The primary efficacy analysis will be performed using a 2-way analysisof covariance (ANCOVA) model with treatment as a factor and baseline TGvalue as a covariate. The least-squares mean, standard error, and2-tailed 95% confidence interval for each treatment group and for eachcomparison will be estimated. The same 2-way ANCOVA model will be usedfor the analysis of secondary efficacy variables.

The primary analysis will be repeated for the per-protocol population toconfirm the robustness of the results for the intent-to-treatpopulation.

Non-inferiority tests for percent change from baseline in LDL-C will beperformed between >96% E-EPA doses and placebo using a non-inferioritymargin of 6% and a significant level at 0.05.

For the following key secondary efficacy parameters, treatment groupswill be compared using Dunnett's test to control the Type 1 error rate:TC, LDL-C, HDL-C, non-HDL-C, VLDL-C, Lp-PLA₂, and apo B. For theremaining secondary efficacy parameters, Dunnett's test will not be usedand the ANCOVA output will be considered descriptive.

The evaluation of safety will be based primarily on the frequency ofadverse events, clinical laboratory assessments, vital signs, and12-lead ECGs. The primary efficacy variable is the percent change infasting TG levels from baseline to Week 12. A sample size of 194completed patients per treatment group will provide 90.6% power todetect a difference of 15% between >96% E-EPA and placebo in percentchange from baseline in fasting TG levels, assuming a standard deviationof 45% in TG measurements and a significance level of p<0.05.

Previous data on fasting LDL-C show a difference in percent change frombaseline of 2.2%, with a standard deviation of 15%, between study drugand placebo. A sample size of 194 completed patients per treatment groupwill provide 80% power to demonstrate non-inferiority (p<0.05,one-sided) of the LDL-C response between >96% E-EPA 4 g daily andplacebo, within a 6% margin. To accommodate a 10% drop-out rate fromrandomization to completion of the double-blind treatment period, atotal of 648 randomized patients is planned (216 patients per treatmentgroup).

Example 2. Ultra-Pure EPA Reduces CVD-Related Parameters in PatientsHaving hsCRP Levels ≥2.0 Mg/L

CVD-related (e.g., atherogenic) parameters were analyzed in a post-hocstudy of 246 patients from the multi-center, placebo-controlled,randomized, double-blind, 12-week study described in Example 1 (i.e.,the ANCHOR trial) who have a baseline hsCRP at least 2.0 mg/L.

Subjects

126 patients were randomized to EPA-A 4 g/day and 120 patients wererandomized to placebo (intent-to-treat “ITT” population). The ITTpopulation was defined as all randomized patients who had a baselinetriglyceride measurement, received at least 1 dose of the study drug,and had at least one post-randomization efficacy measurement performed.In total, atherogenic parameters were analyzed in 246 patients, 55% ofwhich were male, 98% of which were white, and 75% of which had diabetes.The mean age of the 246 patients was 61 years old. Table 7 shows thatbaseline characteristics of the placebo and 4 g per day EPA-E treatmentgroups were similar.

Assessments

The primary objective of this post-hoc study, which is based on an ITTpopulation was to determine the median difference in percent change infasting plasma triglycerides from baseline to week 12 between the groupadministered 4 g per day of EPA-E (e.g., >96% E-EPA) compared to theplacebo group. Additional efficacy assessments included the mediandifference in percent change from baseline to week 12 between the 4 gper day EPA-E and placebo recipients in plasma TC, LDL-C, HDL-C, VLDL-C,VLDL-TG, RLP-C, non-HDL-C, Apo B, Apo C-III, Lp-PLA₂, hsCRP, ox-LDL,IL-6, and ICAM-1 and in plasma and RBC concentrations of EPA. EPA levelswere measured with liquid chromatography using tandem mass spectroscopymethodology in approximately the first 216 patients in ANCHOR withcomplete sample datasets.

Efficacy Assessments

Additional efficacy assessments included the median difference inpercent change from baseline to week 12 between 4 g per day EPA-E andplacebo recipients in plasma TC, LDL-C, HDL-C, VLDL-C, VLDL-TG, RLP-C,non-HDL-C, Apo B, Apo C-III, Lp-PLA2, hsCRP, ox-LDL, IL-6, and ICAM-1and in plasma and RBC concentrations of EPA. These parameters weremeasured as previously described (LDL-C and RLP-C were not calculatedbut were measured by beta-quantification and immunoseparation assays,respectively).

Safety Assessments

The evaluation of safety included monitoring of treatment-emergentadverse events (TEAEs), defined as any AEs that began after the firstdose of study medication or that worsened in severity during thedouble-blind treatment period. TEAEs reported herein included totalTEAEs and those occurring in 3% or less of patients in any treatmentgroup of the overall ANCHOR study population (i.e., nausea, diarrhea,nasopharyngitis, and arthralgia).

Statistical Analysis

The median difference in percent change from baseline between 4 g perday EPA-E and placebo groups for the primary efficacy variable andadditional assessments were estimated with the Hodges-Lehmann method (Pvalues from the Wilcoxon rank-sum test for treatment comparisons) wheredepartures from normality were observed; for normally distributedparameters, an ANCOVA model was used with least square means (LS) andstandard error (SE). Missing values were imputed using thelast-observation-carried-forward method. For all post hoc analyses, thealpha level for statistical significance was 0.05. Safety analyses werebased on the safety population (all randomized patients who received atleast one dose of the study drug).

Results

Patients.

The ANCHOR trial described above in Example 1 randomized 702statin-treated patients to 4 g per day EPA-E, EPA-E 2 g per day, orplacebo. Median baseline hsCRP for the 4 g per day EPA-E, 2 g/day, andplacebo arms of the full ANCHOR cohort were 2.2, 2.0, and 1.8 mg/L,respectively. The present post hoc subgroup analysis included 246patients from the ANCHOR ITT group with baseline hsCRP ≥2.0 mg/Lrandomized to receive 4 g per day EPA-E (n=126) or placebo (n=120). Fourpatients in the 4 g per day EPA-E group and 6 patients in the placebogroup discontinued study medication. As shown in Table 1, baselinecharacteristics were similar in the 4 g per day EPA-E and placebo groups(Table 1). Median (interquartile range) hsCRP level at baseline was 3.9(3.2) mg/L in the 4 g per day EPA-E group and 4.6 (4.4) mg/L in theplacebo group.

TABLE 1 Baseline Characteristics of Patients from ANCHOR study HavingBaseline hsCRP ≥ 2.0 mg/L Icosapent Ethyl 4 g/day Placebo Variable (n =126) (n = 120) Age, mean (SD), years 60.2 (9.7)   61.0 (9.9)   Sex, n(%) Male 72 (57.1) 64 (53.3) Female 54 (42.9) 56 (46.7) Race, n (%)White 124 (98.4)  116 (96.7)  Body mass index, mean (SD), kg/m² 33.7(5.1)   34.0 (5.2)   Diabetes mellitus, n (%) 93 (73.8) 92 (76.7) hsCRP,median (IQR), mg/L^(†) 3.9 (3.2)   4.6 (4.4)   *There were nosignificant differences in baseline characteristics among treatmentgroups in the total randomized ANCHOR population (hsCRP was anexploratory endpoint and was not tested). ^(†)n = 120 for the 4 g/dayEPA-E group and n = 115 for the placebo group.hsCRP, high-sensitivity C-reactive protein; IQR, interquartile range;ITT, intent-to-treat; SD, standard deviation.

Effects on Lipid and Inflammatory Parameters.

As shown in Table 2 and FIG. 1 the administration of 4 g per day EPA-Esignificantly reduced TG by about 19.9% (P<0.0001), without increasingLDL-C levels, in patients with baseline hsCRP 22.0 mg/L compared toplacebo. The administration of 4 g per day EPA-E also significantlyreduced non-HDL-C, TC, HDL-C, VLDL-C, VLDL-TG, and RLP-C as comparedwith placebo (P values ranging from 0.0125 to <0.0001), withoutincreasing LDL-C (P=0.1162; see Table 2 and FIG. 1). Changes in IL-6 andICAM-1 were small and statistically insignificant when compared withplacebo (see Table 2, FIG. 1). Apolipoproteins such as Apo B and ApoC-III and markers of oxidation and inflammation such as hsCRP, ox-LDL,and Lp-PLA₂ were also significantly decreased in patients administered 4g per day EPA-E as compared with placebo (P values ranging from 0.0213to <0.0001; see Table 2, Table 3, and FIG. 1). Changes in IL-6 and ICM-1were small and statistically insignificant when compared to placebo (SeeTable 2 and FIG. 1).

TABLE 2 Effects of EPA-E 4 g/day on Lipid Parameters, Lipoproteins andMarkers of Oxidation, and Inflammation in Patients From the ANCHOR StudyWith Baseline hsCRP ≥2.0 mg/L Median Difference in % Change FromBaseline, Icosapent Ethyl 4 g/day Placebo Icosapent Change Change Ethyl4 g/day End of From End of From vs Placebo, %, Parameter BaselineTreatment Baseline, % Baseline Treatment Baseline, % P ValueLipids/Lipoproteins TG (mg/dL) (primary efficacy variable) n = 126, 120267.8 220.8 −16.6 267.5 280.5 +5 −19.9 (97.0) (110.0) (32.3) (86.8)(118.8) (44.4) <0.0001 LDL-C (mg/dL) n = 126, 119 81.0 83.5 +2.8 84.088.0 +9.2 −5.0 (23.0) (34.0) (28.9) (24.0) (28.0) (36.1) 0.1162Non-HDL-C (mg/dL) n = 126, 120 128.5 125.0 −4.5 131.5 136.0 +9.7 −12.3(29.0) (41.0) (22.0) (29.0) (42.5) (32.7) <0.0001 TC (mg/dL) n = 126,120 167.0 166.0 −2.6 170.0 182.5 +9.2 −11.1 (37.0) (41.0) (15.8) (34.0)(47.5) (24.0) <0.0001 HDL-C (mg/dL) n = 126, 120 36.5 36.0 −2.1 39.039.0 +5.6 −5.2 (12.0) (13.0) (18.8) (11.0) (15.5) (19.6) 0.0042 VLDL-C(mg/dL) n = 126, 119 45.5 38.0 −11.8 42.0 49.0 +11.1 −21.0 (20.0) (20.0)(49.2) (21.0) (29.0) (63.4) <0.0001 VLDL-TG (mg/dL) n = 126, 119 196.5154.5 −18.7 190.0 205.0 +4.6 −22.9 (92.0) (100.0) (52.4) (95.0) (129.0)(67.2) <0.0001 RLP-C (mg/dL)* n = 43, 40 14.0 10.0 −26.3 14.0 13.0 +3.8−23.0 (5.0) (6.0) (45.0) (6.0) (5.0) (67.8) 0.0125 Apolipoproteins Apo B(mg/dL)^(†) n = 120, 115 93.0 93.5 −0.5 91.0 99.0 +6.5 −7.4 (21.5)(24.5) (15.4) (23.0) (22.0) (26.4) 0.0021 Apo C-III (mg/dL)^(†) n = 114,106 15.4 14.0 −7.7 15.1 16.2 +6.8 −16.0 (5.0) (5.0) (28.1) (4.4) (5.6)(33.4) <0.0001 Markers of Oxidation and Inflammation Ox-LDL (U/L)* n =46, 39 54.1 54.1 −2.9 53.3 62.4 +14.7 −13.7 (14.8) (17.2) (20.9) (16.0)(17.9) (25.6) 0.0020 Lp-PLA₂ (ng/mL) n = 120, 113 184.5 163.0 −13.3192.0 203.0 +7.3 −19.6 (54.5) (59.0) (16.3) (61.0) (74.0) (24.6) <0.0001hsCRP (mg/L) n = 120, 115 3.9 3.7 −10.5 4.6 5.4 +8.0 −17.9 (3.2) (3.6)(70.1) (4.4) (7.2) (77.3) 0.0213 IL-6 (pg/mL) n = 46, 38 3.4 2.8 −1.64.2 4.0 −1.7 −4.2 (2.8) (2.5) (57.9) (2.7) (4.2) (66.6) 0.7463 ICAM-1(ng/mL) N = 46, 38 288.5 300.5 +3.3 304.5 307.5 +2.2 +1.1 (107.0)(116.0) (12.1) (130.0) (149.0) (12.9) 0.5621 Data are presented asmedians (interquartile ranges) for endpoint values. Median differencesin percent changes from baseline vs placebo are Hodges-Lehmann medians.Patient numbers are presented as the number of patients in the 4 g/dayEPA-E group and the placebo group, respectively. *RLP-C and ox-LDL weremeasured in approximately the first 35% of patients randomized in theANCHOR study. ^(†)Apo B and Apo C-III were measured in the subgroup ofall patients with available archived plasma samples from the ANCHORstudy.

Significantly, Table 3 shows that lipoproteins and markers of oxidationand inflammation, including hsCRP (−17.9%; P=0.0213), significantlyimproved in patients administered 4 g per day EPA-E (see also FIG. 1).

TABLE 3 Effects of 4 g per day EPA-E on Lipoproteins and Markers ofOxidation and Inflammation in Patients From the ANCHOR Study WithBaseline hsCRP ≥ 2.0 mg/L Median Difference in % Icosapent Ethyl 4 g/dayPlacebo Change From Baseline for End of Change from End of Change fromIcosapent Ethyl 4 g/day vs. Parameter Baseline Treatment Baseline, %Baseline Treatment Baseline, % Placebo, %, P Lipoproteins Apo B (mg/dl)*93.0 93.5 −0.5 91.0 99.0 6.5 −7.4 n = 120, 115 (21.5) (24.5) (15.4)(23.0) (22.0) (26.4) 0.0021 Apo C-III (mg/gL)* 15.4 14.0 −7.7 15.1 16.26.8 −16.0 n = 114, 106 (5.0) (5.0) (28.1) (4.4) (5.6) (33.4) <0.0001Markers of Oxidation and Inflammation hsCRP (mg/L) 3.9 3.7 −10.5 4.6 5.48.0 17.9 n = 120, 115 (3.2) (3.6) (70.1) (4.4) (7.2) (77.3) 0.0213Ox-LDL (U/L)^(†) 54.1 54.1 −2.9 53.3 62.4 14.7 −13.7 n = 46, 39 (14.8)(17.2) (20.9) (16.0) (17.9) (25.6) 0.0020 Lp-PLA₂ (ng/mL) 184.5 163.0−13.3 192.0 203.0 7.3 −19.6 n = 120, 113 (54.5) (59.0) (16.3) (61.0)(74.0) (24.6) <0.0001 Data are presented as median (interquartile range)for endpoint values. Median differences in percent changes from baselineversus placebo are Hodges-Lehmann Medians. Patient numbers are presentedas icosapent ethyl 4 g/day and placebo, respectively. *Apo B and ApoC-III levels were measured in the subgroup of all patients withavailable archived plasma samples from the ANCHOR study. ^(†)Ox-LDL wasonly measured in approximately the first 35% of patients randomized inthe ANCHOR study.

Plasma and RBC Levels of EPA. Administration of 4 g per day of EPA-Esignificantly increased mean (SD) EPA in RBCs from 11.2 (5.5) μg/mL atbaseline to 67.2 (29.3) μg/mL at week 12 (P<0.0001) as shown in Table 4.In patients, administered 4 g per day of EPA-E, EPA levels increased637% in plasma and 632% in RBCs as compared to those who receivedplacebo (both P<0.0001). Similarly, 4 g per day EPA-E significantlyincreased mean (SD) EPA in plasma from 29.4 (20.4) μg/mL at baseline to179.1 (72.9) μg/mL at week 12, representing a LS mean (SE) increase of632% (P<0.0001) versus placebo.

TABLE 4 Change in EPA Levels in Plasma and RBCs from Baseline Week 12 inPatients From the ANCHOR Study With Baseline hsCRP ≥ 2.0 mg/L ChangeFrom Baseline, LS Icosapent Ethyl 4 g/day Placebo Mean, Change ChangeIcosapent Baseline, End of From Baseline, End of From Ethyl 4 g/day MeanTreatment, Baseline, LS Mean Treatment, Baseline, LS vs Placebo, %,Parameter (SD) Mean (SD) Mean (SE) (SD) Mean (SD) Mean (SE) P ValuePlasma EPA, μg/mL n = 41, 38 29.4 179.1 +157.0 25.5 29.7 +11.9 +637.0(20.4) (72.9) (8.7) (14.8) (13.1) (8.8) <0.0001 RBC EPA, μg/mL n = 39,35 11.2 67.2 +57.6 10.0 9.0 +1.0 +631.6 (5.5) (29.3) (3.4) (6.2) (4.4)(3.5) <0.0001 Patient numbers are presented as the number of patients inthe 4 g/day EPA-E group and the placebo group, respectively. *Includesonly patients with available EPA data at both baseline and week 12; EPAwas measured in approximately the first 216 ANCHOR patients withcomplete sample datasets as prespecified. EPA, eicosapentaeneic acid;hsCRP, high-sensitivity C-reactive protein; ITT, intent-to-treat; LS,least squares; RBC, red blood cell; SD, standard deviation; SE, standarderror.

Safety.

As with the overall ANCHOR population of Example 1 above, the safetyprofile of 4 g per day EPA-E was similar to placebo in this subgroupanalysis of patients with baseline hsCRP ≥2.0 mg/L as shown in Table 5.

TABLE 5 Adverse Events in Patients From the ANCHOR Study With BaselinehsCRP ≥ 2.0 mg/L Patients With TEAEs, n (%) Icosapent Ethyl 4 g/dayPlacebo Preferred Term (n = 131) (n = 124) Any TEAE 60 (45.8) 60 (48.4)Nausea* 3 (2.3) 3 (2.4) Diarrhea* 5 (3.8) 8 (6.5) Nasopharyngitis* 1(0.8) 6 (4.8) Arthralgia*^(†) 3 (2.3) 0 (0.0) *TEAEs occurring in ≥ 3%of all patients in the entire safety population of the ANCHOR study (inany treatment group). ^(†)Most commonly reported adverse event listed inthe EPA-E prescribing information. TEAE, treatment-emergent adverseevent.

TEAEs (irrespective of causality) occurred in 60 of 131 (45.8%) patientsin the 4 per day EPA-E group and in 60 of 124 (48.4%) patients in theplacebo group as shown in Table 5. In this subgroup analysis of patientswith ≥2.0 mg/L, the only TEAE reported in 3% or less of patientsreceiving 4 g per day EPA-E was diarrhea, occurring in 5 (3.8%) patientscompared with 8 (6.5%) patients in the placebo group (see Table 5). Inthe overall ANCHOR population (see Example 1), diarrhea, nausea,nasopharyngitis, and arthralgia were the most commonly reported TEAEs(occurring in >3% of any treatment group), and arthralgia was the onlyof these TEAEs that occurred with higher frequency in the EPA-Etreatment groups than in the placebo group. These TEAEs occurred atsimilar rates in this subgroup analysis of patients with baseline hsCRP≥2.0 mg/L (4 g per day EPA-E vs placebo groups, respectively: nausea[2.3% vs 2.4%], nasopharyngitis [0.8% vs 4.8%], and arthralgia [2.3% vs0%]) (see Table 5). One patient each in the 4 g per day EPA-E andplacebo groups discontinued due to an AE (4 g per day EPA-E:subarachnoid hemorrhage, ruptured cerebral aneurysm, and brain edema;placebo: headache), none of which were considered related to study drugby the treating investigator.

CONCLUSION

In summary, this study demonstrated that without increasing LDL-C, EPA-Esignificantly reduced TG (−20%; P<0.0001), non-high-density lipoproteincholesterol (−12.3%; P<0.0001), total cholesterol (−11.1%; P<0.0001),high-density lipoprotein cholesterol (−5.2%; P=0.0042), very-low-densitylipoprotein cholesterol (−21.0%; P<0.0001), very-low-density lipoproteinTG (−22.9%; P<0.0001), remnant lipoprotein cholesterol (−23.0%;P=0.0125), apolipoprotein B (−7.4%; P=0.0021), apolipoprotein C-III(−16%; P<0.0001), oxidized LDL (−13.7%; P=0.0020),lipoprotein-associated phospholipase A2 (−19.6%; P<0.0001), and hsCRP(−17.9%; P=0.0213) vs placebo, while IL-6 and ICAM-1 were notsignificantly changed. EPA levels increased with 4 g per day EPA-E+637%in plasma and +632% in RBCs vs placebo (both P<0.0001). EPA-E exhibiteda safety profile similar to placebo.

Accordingly, this post hoc analysis of patients with hsCRP ≥2.0 mg/L atbaseline suggests icosapent ethyl 4 g per day significantly reduces TGand other atherogenic and inflammatory parameters without increasingLDL-C versus placebo in statin-treated patients with TG between 200-499mg/dL. The results in this elevated hsCRP subgroup are generally similarto those in the entire ANCHOR population. Compared with placebo,icosapent ethyl 4 g/day significantly reduced median TG from baseline by19.9% in the subgroup of patients with hsCRP 22.0 mg/L at baselinecompared with 21.5% in the entire ANCHOR population (all P<0.0001).Icosapent ethyl 4 g/day has demonstrated similar improvement in TG andother atherogenic/inflammatory parameters in other patientsubpopulations of the ANCHOR study, including patients with diabetesmellitus, women, patients with chronic kidney disease, and women withdiabetes mellitus. Importantly, similar reductions in hsCRP have notbeen observed in trials evaluating mixtures of EPA and DHA in patientswith high or very high TG.

Further, in the recently completed Reduction of Cardiovascular Eventswith Icosapent Ethyl-Intervention Trial (REDUCE-IT Clinical Trial),icosapent ethyl 4 g per day resulted in a significant reduction in majoradverse CV events compared with placebo (hazard ratio [HR], 0.75; 95%confidence interval [CI], 0.68-0.83; P<0.001) over a median follow-uptime of 4.9 years. In REDUCE-IT, Log hsCRP decreased by a median of21.8% (P<0.0001) from baseline to the last study visit in the icosapentethyl 4 g per day group and by 0.0% (P=0.9203) in the placebo group,resulting in a 22.5% median between-group decrease for icosapent ethyl 4g/day versus placebo (P<0.0001) at year 2. It is contemplated that thereductions in hsCRP levels may have contributed to the observed CV riskreduction.

Example 3. Improvement of Cognitive Performance in Subjects withAge-Associated Memory Impairment

A single-center, 6-week, double-blind, randomizes, parallel-group,placebo-controlled, dose-ranging pilot study was performed to evaluatethe efficacy, tolerability, and safety of >96% ethyl-EPA in subjectswith subjective and objective memory impairment according to generallyaccepted criteria for Age-Associated Memory Impairment (“AAMI”). Theprimary objective of the study was to determine the effect of >96%ethyl-EPA 1 g, 2 g, and 4 g daily, compared to placebo, on cognitiveperformance in subjects with AAMI.

The secondary objectives of this study were the following:

-   -   1. To determine the effect of >96% E-EPA on the following tests        in the computerized cognitive battery:        -   Continuity of attention tasks;        -   Quality of working memory tasks;        -   Quality of episodic memory tasks; and        -   Speed of attention tasks;    -   2. To determine the safety and tolerability of >96% E-EPA from        routine clinical laboratory tests, adverse events (“AE”)        monitoring, and vital signs; and    -   3. To determine the potential dose-effect relationship of >96%        E-EPA on the cognitive endpoints by measurement of essential        fatty acids in plasma and RBC membranes.

The population for this study was men and women between ages 50 and 70with self-reported complaints of memory loss, subjective and objectivecognitive impairment with a score of at least one standard deviationbelow that of the mean for age-matched elderly population as determinedby the total score of between 13 and 20 from the Paired AssociatedLearning (“PAL”) subset of the Wechsler Memory Scale, evidence ofadequate intellectual function as determined by a scaled score of atleast 9 (raw score of at least 32) on the Vocabulary subtest of theWechsler Adult Intelligence Scale and absence of dementia as determinedby a score of 24 or higher on the Mini-Mental State Examination(“MMSE”).

Potential subjects were excluded based on the following exclusioncriteria:

-   -   Unlikely or unable to comply with investigational medication        dosing requirements;    -   Diagnosis of major depressive disorder, Alzheimer's or vascular        dementia as defined according to the Mini International        Neuropsychiatric Interview (“MINI”)/Diagnostic and Statistical        Manual of Mental Disorders (4th edition) Text Revision (“TR”)        criteria;    -   Past or current history of:        -   a neurological or psychiatric disorder that could have            affected cognitive function;        -   inflammatory gastrointestinal disease such as Crohn's            Disease or ulcerative colitis;        -   cancer other than basal cell carcinoma;        -   clinically significant cardiac abnormality as measured by            12-lead ECG;    -   Any other medical condition or intercurrent illness not        adequately controlled, which, in the opinion of the study        investigator, may have put the subject at risk when        participating in the study or may have influenced the results of        the study or affected the subject's ability to take part in the        study;    -   Clinically significant abnormal screening results (e.g.,        haematology, biochemistry) on screening or vital signs that fell        outside the normal range for this population, which in the        opinion of the study investigator affected the subject's        suitability for the study;    -   Changes to prescribed medication for a medical condition within        4 weeks of the baseline visit;    -   Omega-3 supplementation within 4 weeks of the baseline visit or        during the study treatment period;    -   Currently taking anticoagulants or daily dose of aspirin greater        than 325 mg.    -   Cough or flu remedies containing opiates or antihistamines        within 2 weeks of the baseline visit or during the 6-week        treatment period; and    -   Known allergy to any ingredients of the study drug or placebo.

Ninety-four subjects were randomized into one of six groups: 1 g E-EPAdaily (n=23), 2 g E-EPA daily (n=24), 4 g E-EPA daily (n=24), 1 gplacebo daily (n=7), 2 g placebo daily (n=8), and 4 g placebo daily(n=8). E-EPA was provided as 500 mg soft gel capsules containing >96%E-EPA and 0.2% dl-α-tocopherol as an antioxidant. Placebo capsulescontained 467 mg of liquid paraffin and 0.2% dl-α-tocopherol. Ninety-onesubjects completed the study. Two subjects in the 2 g E-EPA group andone subject in the 2 g placebo group discontinued the study.

The study consisted of a screening visit, a training visit, and fourstudy visits. At the screening visit, subjects' eligibility wasdetermined through cognitive tests (verbal paired associated learning[PAL] subscale, vocabulary subtest, Memory Assessment ClinicsQuestionnaire [MAC-Q], mini mental state evaluation [MMSE] and MINI[mini international neuropsychiatric interview; sections 1 and 2 ofDiagnostic and Statistical Manual of Mental Disorders, 4th Edition(DSM-IV) plus dysthymia]), haematology, clinical chemistry and 12-leadelectrocardiogram (ECG). At the training visit, subjects were shown howto use the CDR computerized system. Subjects took study drug for 6 weeksand on Days 0, 14, 28 and 42, subjects underwent the CDR cognitive testbattery.

At screening cognitive testing and suitability for the study wereassessed using the Verbal Paired Associates 1 (Wechsler Memory Scale),Vocabulary Subtest of the WAIS, MAC-Q, MMSE and MINI (DSM-IV Sections 1and 2 plus Dysthymia).

A selection of tasks from the CDR computerized cognitive assessmentsystem were administered at Visit 2 (training visit), Visit 3(baseline), Visit 4 (Day 14), Visit 5 (Day 28) and Visit 6 (Day 42).Parallel forms of the tests were presented at each testing session. Alltasks were computer-controlled, the information presented on highresolution monitors, and the responses recorded via a response modelcontaining two buttons: one marked ‘no’ and the other ‘yes’. Five CDRcomposite scores were used as the primary/secondary outcome variables.The task titles were:

-   -   Word Presentation    -   Immediate Word Recall    -   Picture Presentation    -   Simple Reaction Time    -   Digit Vigilance    -   Choice Reaction Time    -   Spatial Working Memory    -   Numeric Working Memory    -   Delayed Word Recall    -   Word Recognition    -   Picture Recognition    -   Bond-Lader Visual Analogue Scales of Mood and Alertness    -   Screen, Using the Computer Mouse

To ensure consistency of approach, full training on the cognitive testsand CDR test battery was provided to study site staff and studysubjects. The results of each variable were automatically recorded usingthe machine interface developed by CDR.

Blood samples (10 mL) were collected at Visit 1 (screening) and atVisits 4, 5 and 6. Analysis was performed by MSR Lipid Analysis,Scottish Crop Research Institute, Dundee, UK. The screening sample actedas baseline for the EFA measurements. Lipid was extracted from plasma,serum and RBC suspensions and converted into fatty acid methyl esterswhich were analyzed by gas chromatography to give fatty acid profiles asmicrograms fatty acid per gram of sample (μgFA/g) and normalized areapercent.

All randomized subjects with at least 1 visit post-baseline wereincluded in the Intent to Treat (“ITT”) population, regardless oftreatment actually received.

All randomized subjects that completed the study, excluding significantprotocol deviators, were defined as the Safety Per Protocol population.An Efficacy Per Protocol population was based on the Efficacycompleters. The intercept of the Safety and Efficacy Per Protocolpopulations defined the Study Per Protocol Population.

All randomized subjects that received at least 1 dose of studymedication were included in the Safety Population.

Summary statistics were provided for the ITT and Study Per ProtocolPopulations separately for all composite scores, major and supportivevariables. Summary statistics were performed for both the unadjusted anddifference from baseline data (i.e. the difference from the time matchedpredose assessments on Day 0). Summary statistics were calculated bytreatment, day and time-point. The summary statistics comprised n, mean,median, SD, standard error of mean (“SEM”), minimum and maximum values.

Difference from baseline data for each major variable was evaluated byan Analysis of Covariance (“ANCOVA”) using SAS® PROC MIXED Version 8.2.Fixed effects for treatment, day, time point, treatment by day,treatment by time point, treatment by day by time-point were fitted.Subject within treatment was fitted as a repeated effect using therepeated statement. The compound symmetry covariance structure was used.Subjects' time-matched predose assessments on Day 0 were used as acovariate in the analysis.

Least squares means (LS means) were calculated for treatment by day,treatment by time-point and treatment by day by time-point interaction.This formal analysis was conducted for the ITT and Study PP Populationsseparately.

Safety evaluations were based on the safety population. Safety andtolerability were assessed in terms of AEs, vital signs, 12-lead ECG,clinical laboratory data, medical history, and study drug compliance.Safety and tolerability data were presented by treatment group.

RBC and plasma EFA data were collected at baseline, Day 14, 28 and 42and summarized by visit for each treatment group. Change from baselineand percent change from baseline were also summarized. ANCOVA comparisonof ethyl-EPA dose groups and ethyl-EPA versus placebo was performed.

Efficacy Results.

All CDR cognitive test battery analyses were completed for the ITT andStudy PP analysis populations.

For the Intent-to-Treat Analysis for Power of Attention, there was nostatistically significant effect of treatment, nor any treatment by day,treatment by time-point or treatment by day by time-point interactions.There was no LS mean difference between active treatment and placebo atany time-point.

For the contributing subtasks Simple Reaction Time and Digit VigilanceSpeed, there were no statistically significant effects of treatment, norany treatment by day, treatment by time-point or treatment by day bytime-point interactions. For the subtask measure Choice Reaction Time,there was a statistically significant treatment by day interaction(p=0.011).

For the Study Per-Protocol Power of Attention, there was nostatistically significant effect of treatment, nor any treatment by day,treatment by time-point or treatment by day by time-point interactions.There was no difference between active treatment and placebo at anytime-point.

For the subtasks Simple Reaction Time and Digit Vigilance Speed, therewere no statistically significant effects of treatment, nor anytreatment by day, treatment by time-point or treatment by day bytime-point interactions. For the subtask measure, Choice Reaction Time,there was a statistically significant treatment by day interaction(p=0.013).

The Intent-to-Treat Continuity of Attention and the contributing subtaskDigit Vigilance Targets Detected tests showed no statisticallysignificant effect of treatment, nor any treatment by day, treatment bytime-point or treatment by day by time-point interactions.

For the Study Per Protocol Continuity of Attention test, there was nostatistically significant effect of treatment, nor any treatment by day,treatment by time-point or treatment by day by time-point interactions.

For the subtask Digit Vigilance Targets Detected, there was astatistically significant treatment by time-point interaction (p=0.040).

For the Intent-to-Treat Quality of Working Memory test, there was astatistically significant treatment by day interaction (p=0.019).

For the contributing subtask Spatial Working Memory Sensitivity Index,there was a statistically significant treatment by day interaction(p=0.015).

For Numeric Working Memory Sensitivity Index, there was a statisticaltrend for a treatment by day interaction (p=0.089).

For the Study Per-Protocol Quality of Working Memory test, there was astatistically significant treatment by day interaction (p=0.021).

For the contributing subtask Spatial Working Memory Sensitivity Index,there was a statistically significant treatment by day interaction(p=0.014).

For the Intent-to-Treat Quality of Episodic Secondary Memory test, therewas no statistically significant effect of treatment, nor any treatmentby day, treatment by time-point or treatment by day by time-pointinteractions. The LS mean differences showed overall statisticallysignificant decreases versus placebo for ethyl-EPA 1 g and 2 g (p=0.040and p=0.035, respectively).

For the contributing subtasks Immediate and Delayed Word RecallAccuracies and for Word and Picture Recognition Sensitivity Indices,there were no statistically significant effects of treatment ortreatment by day, treatment by time-point or treatment by day bytime-point interactions. For Immediate Word Recall Accuracy, the LS meandifferences showed statistically significant decreases for 1 g on Day 14(p=0.028) and for 2 g on Day 28 (p=0.017). There were statisticallysignificant decreases versus placebo for 1 g and 2 g at AM 1 (p=0.040and p=0.028, respectively). There were statistically significantdecreases for ethyl-EPA 1 g versus placebo on Day 14 at PM 2 (p=0.020)and for 2 g on Day 28 at AM 1 (p=0.006). For Word RecognitionSensitivity Index, the LS mean differences showed statisticallysignificant decreases for ethyl-EPA 1 g on Day 28 (p=0.024) and for 4 gon Day 42 (p=0.038) versus placebo. There was a statisticallysignificant decrease for 4 g at PM 2 (p=0.045) and a statisticallysignificant decrease for 4 g versus placebo on Day 28 at PM 2 (p=0.030).For Picture Recognition Sensitivity Index, the LS mean differencesshowed statistically significant decrease for 1 g versus placebo on Day28 at AM 2 (p=0.017) and at PM 2 (p=0.040). For the Study Per-ProtocolQuality of Episodic Secondary Memory test, there were no statisticallysignificant effects of treatment, nor treatment by day, treatment bytime-point or treatment by day by time-point interactions. The LS meandifferences showed overall statistically significant decreases versusplacebo for 1 g and 2 g (p=0.043 and p=0.036, respectively).

For the contributing subtasks Immediate and Delayed Word RecallAccuracies and for Word and Picture Recognition Sensitivity Indices,there were no statistically significant effects of treatment ortreatment by day, treatment by time-point or treatment by day bytime-point interactions. For Immediate Word Recall Accuracy, the LS meandifferences to placebo showed statistically significant decreases forethyl-EPA 1 g on Day 14 (p=0.024) and for 2 g on Day 28 (p=0.017). Therewere statistically significant decreases for 1 g and 2 g at AM 1(p=0.038 and p=0.029, respectively) and for 1 g at AM 2 (p=0.048). Therewere statistically significant decreases for 1 g versus placebo on Day14 at PM 2 (p=0.019) and for 2 g on Day 28 at AM 1 (p=0.006).

For Word Recognition Sensitivity Index, the LS mean differences toplacebo showed statistically significant decreases for 4 g on Day 42(p=0.038) and for 1 g on Day 28 (p=0.027).

For Picture Recognition Sensitivity Index, the LS mean differencesshowed statistically significant decreases versus placebo for 1 g on Day28 at AM 2 (p=0.020) and PM 2 (p=0.026).

For Intent-to-Treat Speed of Memory and the contributing subtasksSpatial and Numeric Working Memory Speeds and Word, and PictureRecognition Speeds, there were no statistically significant effects oftreatment, nor treatment by day, treatment by time-point or treatment byday by time-point interactions. For Spatial Working Memory Speed, the LSmean differences showed a statistically significant benefit versusplacebo for ethyl-EPA 4 g on Day 14 at PM 1 (p=0.048) and a trend for abenefit for 4 g on Day 42 at AM 1 (p=0.061). For Picture RecognitionSpeed, there were trends for benefits versus placebo for 1 g on Day 14at AM 2 (p=0.084) and on Day 28 at AM 1 (p=0.085).

For Study Per-Protocol Speed of Memory and the contributing subtasksSpatial and Numeric Working Memory Speed and Word, and PictureRecognition Speed, there were no statistical significant effects oftreatment, nor any treatment by day, treatment by time-point ortreatment by day by time-point interactions.

For Intent-to-Treat Self-Rated Alertness, there was no statisticallysignificant effect of treatment, nor any treatment by day, treatment bytime-point or treatment by day by time-point interactions. The LS meandifferences showed a statistically significant decrease in ratings forethyl-EPA 2 g on Day 28 (p=0.047) versus placebo. There was astatistically significant decrease in ratings versus placebo for 2 g onDay 28 at AM 2 (p=0.041). For Study Per-Protocol Self-Rated Alertness,there was no statistically significant effect of treatment, nor anytreatment by day, treatment by time-point or treatment by day bytime-point interactions. The LS mean differences showed a statisticallysignificant decrease in ratings for ethyl-EPA 2 g on Day 28 (p=0.035)versus placebo. There was a statistically significant decrease inratings versus placebo for 2 g on Day 28 at AM 2 (p=0.033).

For Intent-to-Treat Self-Rated Contentment, there was a statisticallysignificant treatment by day interaction (p<0.001). The LS meandifference to placebo showed no statistically significant effects. ForStudy Per-Protocol Self-Rated Contentment, there was a statisticallysignificant treatment by day interaction (p<0.001). The LS meandifference to placebo showed no statistically significant effects.

For Intent-to-Treat Self-Rated Calmness, there was no statisticallysignificant effect of treatment, nor any treatment by day, treatment bytime-point or treatment by day by time-point interactions. For StudyPer-Protocol Self-Rated Calmness, there was no statistically significanteffect of treatment, nor any treatment by day, treatment by time-pointor treatment by day by time-point interactions. The LS mean differencesshowed a statistical trend for an increase in ratings versus placebo forethyl-EPA 4 g on Day 42 at PM 1 (p=0.071).

A post-hoc analysis compared the individual placebo groups (1 g, 2 g and4 g paraffin oil) with the corresponding ethyl-EPA doses.

The pattern of data provided evidence that ethyl-EPA 4 g might improvespeed in the attention based measures. For Power of Attention, there wasan overall benefit versus the corresponding placebo for 4 g on Day 42.The subtask Simple Reaction Time showed improvements in performance for4 g at PM 2 collapsed across days and at several time-points on Days 14and 42. The improvements for 4 g were most pronounced in the ChoiceReaction Time task, where there was an overall benefit versuscorresponding placebo for 4 g, reflecting a benefit for 4 g over placeboon all study days. The pattern of improvement in performance throughoutthe assessment days was quite convincing as the improvements began onDay 14 with improvements seen at 2 time points, whereas on Day 42ethyl-EPA 4 g was superior to placebo at every time point.

For Continuity of Attention, there were isolated declines orimprovements in performance, but there was no general pattern of effectsand it was considered unlikely these differences were due to the studycompound. For Quality of Working Memory and in the subtask measureNumeric Working Memory Sensitivity Index, there were, as in the originalanalyses, only isolated improvements and declines in performance thatwere most likely not treatment-related. However, for Spatial WorkingMemory Sensitivity Index, there was an overall benefit for ethyl-EPA 4 gover placebo on Day 42 in the Study PP Population, which corresponds tothe improvements seen for the attention based measures.

For Quality of Episodic Secondary Memory and contributing subtasks,there were a number of decreases for ethyl-EPA that could be explainedby the pre-existing differences in performance between the placebo andactive treatment groups which was seen in the original analyses. Incontrast to the original analysis, the subtask measures of Speed ofMemory showed some signs of improvement in performance for activetreatment, mostly for 1 g versus placebo. For Self-rated Alertness andSelf-rated Contentment, the 1 g dose showed decreases in ratings on Days14 and 28. However, these decreases were not correlated with a declinein performance in the CDR attention tasks. As with the original plannedanalysis, there were no differences between active treatment and placeboin Self-Rated Calmness.

Safety Results.

Subjects who used less than 80% of the prescribed dose were to beconsidered non-compliant; other than those subjects who withdrew forother reasons only 1 subject fell into this category and was withdrawn.

Overall, 139 treatment emergent AEs (“TEAEs”) were reported by 62(66.0%) of subjects during the study. Most TEAEs were considered mild inseverity and unrelated to study drug. More TEAEs were reported for theethyl-EPA treatment groups (105 events) compared to the placebotreatment groups (34 events). One SAE was reported for the ethyl EPA 2 gtreatment group and 3 subjects discontinued due to TEAEs: 2 subjectsfrom the ethyl-EPA 2 g treatment group (the primary reason fordiscontinuation for 1 of these subjects was non-compliance), and 1subject from the placebo 2 g treatment group.

There were no deaths during the study. No TEAEs were Definitely Relatedto the study drug. One subject receiving 1 g ethyl-EPA experiencednausea that was Probably Related to the study drug. Another subjectreceiving 4 g ethyl-EPA experienced diarrhea that was Probably Relatedto the study drug; another subject receiving 2 g placebo alsoexperienced diarrhea that was Probably Related to the study drug. Fivesubjects experienced nausea that was Possibly Related to the study drug;two were in the 1 g ethyl-EPA cohort; one was in the 2 g ethyl-EPAcohort; two were in the 4 g ethyl-EPA cohort. One subject receiving 2 gplacebo experienced headache that was Possibly Related to the studydrug. All other TEAEs were Not Related or Unlikely Related to the studydrug, and included nasopharyngitis (n=3), cystitis (n=2), cough (n=7),toothache (n=2), pharyngolaryngeal pain (n=2), back pain (n=2),pollakiuria (n=2), influenza-like illness (n=2), headache (n=15),diarrhea (n=2), and nausea (n=1).

One subject with a history of transient ischemic attack, hypertensionand osteoarthritis of the hand and osteopenia receiving 2 g ethyl-EPAexperienced worsening epigastric chest pain 17 days after the start ofthe study and 9 days after the last dose of the study drug. A plannedendoscopy revealed oesophagitis and a small hiatus hernia. The subjectwas treated with omeprazole, which settled her symptoms. The subject hadtaken felodipine, rosuvastatin, aspirin, glucosamine, and quinine within14 days of the onset of her symptoms. The study investigator determinedthat her symptoms were unrelated to the study drug and withdrew thesubject from the study. No other Serious Adverse Events occurred duringthe study.

Essential fatty acid parameters in plasma and RBCs was measured atbaseline and on Day 14, 28 and 48 (shown in Tables 6-11). Notablechanges for these parameters occurred in the ethyl-EPA treatment groupsat Days 14, 28 and 42 compared to placebo. EPA, DPAn-3 and EPA/AA ratiovalues increased substantially from baseline, in plasma and RBC, to Day42 for the ethyl-EPA 1, 2 and 4 g treatment groups, but remained similarto baseline in the placebo treatment groups. AA, DHA and DGLA valuesdecreased substantially from baseline, in plasma and RBC, to day 42 forthe ethyl EPA 1, 2 and 4 g treatment groups, but remained similar tobaseline in the placebo treatment groups. The difference in EPA, AA (RBConly), DPAn-3, DGLA (1 g only for plasma) and EPA/AA ratio levels in theplasma and RBC were significantly (LS means, p≤0.05) different for theethyl-EPA 4 g treatment group compared to the ethyl-EPA 1 g and 2 gtreatment groups.

TABLE 6 EFA Parameter EPA (Plasma and RBC) Mean change from Baseline toDays 14, 28 and 42. Ethyl-EPA Placebo EFA Parameter 1 g 2 g 4 g 1 g 2 g4 g (μg/g) (N = 23) (N = 24) (N = 24) (N = 7) (N = 8) (N = 8) PlasmaBaseline: n 23 24 24 7 8 3 Mean (SD) 48.3 (31.03)  44.9 (25.01)  49.1(17.23) 47.5 (26.41)  42.1 (16.18) 42.5 (11.86) Day 14: n 23 22 24 7 7 8Mean (SD) 61.2 (26.61) 124.6 (42.25) 207.7 (57.05)  1.6 (24.69)  −1.2(19.82) 21.9 (32.91) Day 28: n 22 22 24 7 7 8 Mean (SD) 60.3 (36.03)142.2 (46.23) 215.2 (58.68)  6.5 (15.46)  1.6 (13.64)  1.3 (14.03) Day42: n 23 22 24 7 7 3 Mean (SD) 62.0 (39.43) 133.4 (43.34) 204.6 (80.69)11.9 (26.34)  0.4 (21.18)  4.4 (23.32) 1 or 2 g versus 4 g LS Mean−111.8   −60.9  — — — — CI −123.6, −100   −7.27, −49.0 — — — — p-value   <0.001    <0.001 RBC Baseline: n 23 24 24 7 7 8 Mean (SD) 19.8(10.85) 18.9 (8.91) 19.8 (5.28) 20.4 (5.77)  19.3 (6.58) 17.2 (4.94) Day 14: n 23 22 24 7 7 3 Mean (SD) 12.3 (7.39)  26.9 (9.15)  39.5(13.16) −0.5 (6.32)   0.0 (7.17) 2.6 (6.73) Day 28: n 22 22 24 7 7 8Mean (SD) 14.5 (10.47)  32.9 (10.11)  50.2 (15.82) 1.5 (4.16)  0.0(7.06) 0.6 (4.42) Day 42: n 23 22 24 7 7 8 Mean (SD) 17.6 (11.89)  38.3(12.46)  52.5 (20.56) −0.2 (5.90)   1.0 (8.01) −0.2 (6.97)  1 or 2 gversus 4 g LS Mean −24.4  −11.8  — — — — CI −27.6, −21.2 −15.0, −8.6  —— — — p-value    <0.001    <0.001

TABLE 7 EFA Parameter AA (Plasma and RBC) Mean change from Baseline toDays 14, 28 and 42. Ethyl-EPA Placebo EFA Parameter 1 g 2 g 4 g 1 g 2 g4 g (μg/g) (N = 23) (N = 24) (N = 24) (N = 7) (N = 8) (N = 8) PlasmaBaseline: n 23 24 24 7 8 8 Mean (SD) 202.5 (44.40) 227.3 (42.26) 220.9(42.80) 210.7 (35.68)  191.6 (28.24) 248.0 (53.52) Day 14: n 23 22 24 77 8 Mean (SD)  −9.7 (22.20) −13.9 (22.13) −27.2 (28.39) 0.8 (40.00)−14.4 (19.45)  −5.9 (25.00) Day 28: n 22 22 24 7 7 8 Mean (SD) −11.3(28.13)  21.6 (28.32) −43.7 (32.24) 3.8 (28.11)  −7.4 (23.72) −16.4(31.42) Day 42: n 23 22 24 7 7 8 Mean (SD)  −8.7 (31.35) −27.3 (26.76)−48.3 (22.20) 8.2 (20.30) −11.5 (20.88) −11.0 (25.82) 1 or 2 g versus 4g LS Mean   4.2   15.6 — — — — CI −8.0, 16.4 3.4, 27.8 — — — — p-value   0.496    0.013 RBC Baseline: n 23 24 24 7 8 8 Mean (SD) 171.2 (19.79)172.8 (22.79) 171.0 (25.17) 176.4 (17.65)  152.8 (17.36) 180.4 (23.68)Day 14: n 23 22 24 7 7 8 Mean (SD)  −8.1 (21.95)  −3.1 (25.84) −15.7(26.76) −8.5 (22.75)   3.0 (18.20)  −8.1 (27.53) Day 28: n 22 22 24 7 78 Mean (SD) −17.0 (20.69) −14.1 (26.89) −22.8 (29.56) 5.2 (22.95)  −2.6(17.78)  −8.2 (26.89) Day 42: n 23 22 24 7 7 8 Mean (SD) −14.2 (27.69)−18.8 (25.62) −34.4 (31.44) −9.8 (21.59)   9.7 (16.58) −10.6 (33.49) 1or 2 g versus 4 g LS Mean   8.4   9.8 — — — — CI  2.0, 14.9 3.3, 16.2 —— — — p-value    0.010    0.003

TABLE 8 EFA Parameter DHA (Plasma and RBC) Mean change from Baseline toDays 14, 28 and 42. Ethyl-EPA Placebo EFA Parameter 1 g 2 g 4 g 1 g 2 g4 g (μg/g) (N = 23) (N = 24) (N = 24) (N = 7) (N = 8) (N = 8) PlasmaBaseline: n 23 24 24 7 8 8 Mean (SD) 73.1 (30.43) 75.1 (24.02)   78.8(19.00) 73.7 (14.21) 73.3 (27.74) 76.7 (15.68) Day 14: n 23 22 24 7 7 8Mean (SD) −6.4 (13.30) −5.4 (14.29) −10.3 (13.35)  0.4 (18.36) −0.8(14.28) 13.8 (21.05) Day 28: n 22 22 24 7 7 8 Mean (SD) −6.6 (15.53)−8.1 (15.82) −13.5 (14.10)  4.7 (16.31) −0.6 (8.29)   6.0 (17.36) Day42: n 23 22 24 7 7 8 Mean (SD) −5.4 (18.17) −6.0 (16.69) −13.8 (15.31)11.8 (21.27)  0.8 (17.57)  6.2 (13.40) 1 or 2 g versus 4 g LS Mean  −0.8   1.5 — — — — CI −7.3, 5.7 −5.0, 8.1 — — — — p-value    0.810   0.644 RBC Baseline: n 23 24 24 7 8 8 Mean (SD) 66.5 (18.65) 64.8(17.65)   68.3 (14.24) 71.1 (7.48)  66.0 (15.90) 66.2 (15.83) Day 14: n23 22 24 7 7 8 Mean (SD) −4.6 (9.76)  −2.0 (9.46)  −6.9 (9.13) −5.5(11.93) −0.2 (12.39) −0.4 (12.50) Day 28: n 22 22 24 7 7 8 Mean (SD)−6.4 (11.57) −6.2 (9.34)   −8.7 (11.63)  0.6 (12.86) −0.3 (11.29)  1.1(12.54) Day 42: n 23 22 24 7 7 8 Mean (SD) −7.0 (12.20) −6.3 (9.42) −13.8 (13.76) −4.1 (12.02)  4.6 (12.94) −0.1 (17.63) 1 or 2 g versus 4 gLS Mean   1.0   1.0 — — — — CI −3.5, 5.4 −3.5, 5.5 — — — — p-value   0.674    0.664

TABLE 9 EFA Parameter DPAn-3 (Plasma and RBC) Mean change from Baselineto Days 14, 28 and 42. Ethyl-EPA Placebo EFA Parameter 1 g 2 g 4 g 1 g 2g 4 g (μg/g) (N = 23) (N = 24) (N = 24) (N = 7) (N = 8) (N = 8) PlasmaBaseline: n 23 24 24 7 8 8 Mean (SD) 21.1 (6.62)  19.7 (4.50) 21.7(4.69)  17.9 (5.18) 18.0 (4.39) 19.0 (2.67) Day 14: n 23 22 24 7 7 8Mean (SD) 7.5 (5.11) 17.4 (7.49) 24.5 (11.28) −0.2 (3.13) −1.0 (3.59) 2.2 (4.93) Day 28: n 22 22 24 7 7 8 Mean (SD) 3.9 (5.62) 19.4 (8.48)29.7 (13.23)  1.2 (2.06)  0.6 (3.44)  1.3 (3.40) Day 42: n 23 22 24 7 78 Mean (SD) 11.3 (6.61)  19.3 (8.63) 32.0 (16.01)  2.2 (3.29)  0.1(3.61)  0.8 (6.70) 1 or 2 g versus 4 g LS Mean  −15.1   −9.5 — — — — CI−17.6, −12.7 −12.0, −7.1 — — — — p-value    <0.001    <0.001 RBCBaseline: n 23 24 24 7 8 8 Mean (SD) 34.1 (5.43)  33.2 (4.51) 34.5(4.34) 34.0 (4.27) 33.0 (1.20) 32.4 (2.41) Day 14: n 23 22 24 7 7 8 Mean(SD) 0.9 (5.03)  5.6 (6.28) 5.4 (5.38) −2.8 (4.86) −0.3 (4.96) −0.9(4.74) Day 28: n 22 22 24 7 7 8 Mean (SD) 3.3 (5.42)  9.4 (6.74) 12.4(6.98)   0.1 (4.51) −0.8 (4.03) −0.6 (5.19) Day 42: n 23 22 24 7 7 8Mean (SD) 6.5 (6.19) 13.2 (7.23) 16.2 (10.07) −1.8 (4.64)  2.2 (4.44)−0.9 (6.03) 1 or 2 g versus 4 g LS Mean   −6.2   −2.5 — — — — CI −7.8,−4.7 −4.1, −1.0 — — — — p-value    <0.001    0.002

TABLE 10 EFA Parameter DGLA (Plasma and RBC) Mean change from Baselineto Days 14, 28 and 42. Ethyl-EPA Placebo EFA Parameter 1 g 2 g 4 g 1 g 2g 4 g (μg/g) (N = 23) (N = 24) (N = 24) (N = 7) (N = 8) (N = 8) PlasmaBaseline: n 23 24 24 7 8 8 Mean (SD)  51.2 (15.01)  53.5 (14.12)  57.1(14.73) 51.6 (9.20)  41.6 (10.30) 52.6 (7.74) Day 14: n 23 22 24 7 7 8Mean (SD) −10.4 (10.90) −14.1 (6.88)  −22.9 (9.00)  −4.1 (8.07) −0.0(8.63)  −1.0 (11.58) Day 28: n 22 22 24 7 7 8 Mean (SD) −10.6 (10.23)−16.2 (9.88)  −24.2 (10.73) −4.6 (7.43) −0.6 (5.91)  1.5 (11.78) Day 42:n 23 22 24 7 7 8 Mean (SD) −9.4 (9.41) −17.3 (9.92)  −22.5 (10.87) −3.9(12.90)  0.9 (9.34)  0.8 (11.04) 1 or 2 g versus 4 g LS Mean   3.7   2.5— — — — CI 0.4, 7.0 −0.9, 5.8 — — — — p-value    0.028    0.143 RBCBaseline: n 23 24 24 7 8 8 Mean (SD) 23.0 (5.19) 23.0 (5.76) 24.0 (5.77)22.4 (5.06) 19.7 (5.87) 22.4 (4.91) Day 14: n 23 22 24 7 7 8 Mean (SD)−2.7 (3.82) −2.6 (3.54) −5.3 (4.10) −1.5 (2.08)  0.2 (1.76) −1.8 (4.00)Day 28: n 22 22 24 7 7 8 Mean (SD) −3.8 (3.31) −4.5 (3.58) −7.1 (4.63) 0.2 (3.63) −0.7 (4.06) −0.7 (3.81) Day 42: n 23 22 24 7 7 8 Mean (SD)−3.5 (4.51) −5.3 (3.65) −8.0 (4.98) −1.6 (4.93)  1.9 (3.61) −1.1 (5.31)1 or 2 g versus 4 g LS Mean   1.5   1.5 — — — — CI 0.2, 2.9   0.1, 2.9 —— — — p-value    0.027    0.032

TABLE 11 EFA Parameter EPA/AA (Plasma and RBC) Mean change from Baselineto Days 14, 28 and 42. Ethyl-EPA Placebo EFA Parameter 1 g 2 g 4 g 1 g 2g 4 g (μg/g) (N = 23) (N = 24) (N = 24) (N = 7) (N = 8) (N = 8) PlasmaBaseline: n 23 24 24 7 8 8 Mean (SD) 0.2 (0.14) 0.2 (0.12) 0.2 (0.07)0.2 (0.11) 0.2 (0.10) 0.2 (0.07) Day 14: n 23 22 24 7 7 8 Mean (SD) 0.3(0.4)  0.6 (0.23) 1.1 (0.28) 0.0 (0.09) 0.0 (0.12) 0.1 (0.12) Day 28: n22 22 24 7 7 8 Mean (SD) 0.3 (0.20) 0.8 (0.35) 1.3 (0.42) 0.0 (0.08) 0.0(0.09) 0.0 (0.06) Day 42: n 23 22 24 7 7 8 Mean (SD) 0.3 (0.24) 0.7(0.29) 1.3 (0.45) 0.0 (0.10) 0.0 (0.12) 0.0 (0.08) 1 or 2 g versus 4 gLS Mean   −0.66   −0.41 — — — — CI −0.731, −0.597 −0.475, −0.341 — — — —p-value    <0.001    <0.001 RBC Baseline: n 23 24 24 7 8 8 Mean (SD) 0.1(0.07) 0.1 (0.06) 0.1 (0.04) 0.1 (0.04) 0.1 (0.06) 0.1 (0.03) Day 14: n23 22 24 7 7 8 Mean (SD) 0.1 (0.04) 0.2 (0.04) 0.3 (0.07) 0.0 (0.03)−0.0 (0.05)   0.0 (0.03) Day 28: n 22 22 24 7 7 8 Mean (SD) 0.1 (0.05)0.02 (0.06)  0.4 (0.11) 0.0 (0.01) −0.0 (0.04)   0.0 (0.02) Day 42: n 2322 24 7 7 8 Mean (SD) 0.1 (0.06) 0.3 (0.06) 0.4 (0.14) 0.0 (0.03) −0.0(0.05)   0.0 (0.03) 1 or 2 g versus 4 g LS Mean   −0.18   −0.11 — — — —CI −0.204, −0.162 −0.126, −0.085 — — — — p-value    <0.001    <0.001

I/We claim:
 1. A method of lowering triglycerides in a subject on stablestatin therapy, the method comprising orally administering to thesubject daily a pharmaceutical composition comprising about 4 g of ethyleicosapentaenoate and not more than about 4% docosahexaenoic acid or itsesters, by weight of all fatty acids, for a period of at least about 4weeks, wherein the subject has baseline fasting triglyceride levels ofabout 200 mg/dl to about 500 mg/dl and high-sensitivity C-reactiveprotein (hsCRP) levels of at least about 2 mg/L.
 2. The method of claim1, wherein the subject has mixed dyslipidemia, chronic kidney disease,and/or reduced kidney function.
 3. The method of claim 2, wherein thesubject is determined to have chronic kidney disease if the subject hasan estimated glomerular filtration rate (eGFR) of less than about 60ml/min/1.73 m² for at least about 3 months and/or reduced kidneyfunction if the subject has an eGFR of less than about 90 mL/min/1.73 m²for at least about 3 months.
 4. The method of claim 1, wherein thesubject has coronary heart disease or a coronary heart disease riskequivalent.
 5. The method of claim 3, wherein the subject has alow-density lipoprotein cholesterol (LDL-C) level of about 40 mg/dl toabout 100 mg/dl prior to said administering.
 6. The method of claim 1,wherein LDL-C is not increased in the subject upon 4 weeks of saidadministering.
 7. The method of claim 1, wherein LDL-C is reduced in thesubject upon 4 weeks of said administering.
 8. The method of claim 1,wherein the subject exhibits a reduction in one or more of:non-high-density lipoprotein cholesterol (non-HDL-C), very-low-densitylipoprotein cholesterol (VLDL-C), lipoprotein-associated phospholipaseA2 (Lp-PLA₂), apolipoprotein B (Apo B), total cholesterol (TC),high-density lipoprotein cholesterol (HDL-C), very-low-densitylipoprotein triglyceride (VLDL-TG), remnant particle like-cholesterol(RLP-C), or hsCRP levels.
 9. The method of claim 6, wherein the subjectexhibits a reduction in one or more of: non-HDL-C, VLDL-C, Lp-PLA₂, ApoB, TC, HDL-C, VLDL-TG, RLP-C, and hsCRP levels.
 10. The method of claim1, wherein the subject exhibits an increase in plasma and/or red bloodcell (RBC) levels of EPA.
 11. The method of claim 1, wherein thepharmaceutical composition is present in one or more dosage units.
 12. Amethod of lowering triglycerides in a subject on stable statin therapy,the method comprising orally administering to the subject daily apharmaceutical composition comprising about 4 g of ethyleicosapentaenoate and not more than about 4% docosahexaenoic acid or itsesters, by weight of all fatty acids, for a period of at least about 4weeks, wherein the subject has a baseline fasting triglycerides of about200 mg/dl to about 500 mg/dl, low-density lipoprotein cholesterol(LDL-C) levels of about 40 mg/dl to about 100 mg/dl, andhigh-sensitivity C-reactive protein (hsCRP) levels of at least about 2mg/L.
 13. The method of claim 12, wherein the subject has mixeddyslipidemia, chronic kidney disease, and/or reduced kidney function.14. The method of claim 13, wherein the subject is determined to havechronic kidney disease if the subject has an estimated glomerularfiltration rate (eGFR) of less than about 60 mL/min/1.73 m² for at leastabout 3 months and/or reduced kidney function if the subject has an eGFRof less than about 90 mL/min/1.73 m² for at least about 3 months. 15.The method of claim 1, wherein the subject has coronary heart disease ora coronary heart disease risk equivalent.
 16. The method of claim 12,wherein LDL-C levels are not increased in the subject upon 4 weeks ofsaid administering.
 17. The method of claim 12, wherein LDL-C is reducedin the subject upon 4 weeks of said administering.
 18. The method ofclaim 16, wherein the subject exhibits a reduction in one or more of:(non-high-density lipoprotein cholesterol) non-HDL-C, (very-low-densitylipoprotein cholesterol) VLDL-C, lipoprotein-associated phospholipase A2(Lp-PLA₂), apolipoprotein B (Apo B), total cholesterol (TC),high-density lipoprotein cholesterol (HDL-C), very-low-densitylipoprotein triglyceride (VLDL-TG), remnant particle like-cholesterol(RLP-C), or hsCRP levels.
 19. The method of claim 12, wherein thesubject exhibits an increase in plasma and/or red blood cell (RBC)levels of EPA.
 20. The method of claim 12, wherein the pharmaceuticalcomposition is present in one or more dosage units.